Heparan sulfate/heparin mimetics with anti-chemokine and anti-inflammatory activity

ABSTRACT

The present disclosure provides for methods and compositions comprising a series of synthetic glycopolymers. The disclosure also relates to a kit which is suitable for carrying out the inventive methods.

CROSS-REFERENCE

This application claims the benefit of U.S. Provisional Application No.61/861,627, filed on Aug. 2, 2013, which application is incorporatedherein by reference in its entirety.

STATEMENT AS TO FEDERALLY SPONSORED RESEARCH

This invention was made with government support under Grant No. GM093627awarded by the National Institutes of Health. The government has certainrights in the invention.

BACKGROUND OF THE INVENTION

Glycosaminoglycans, a ubiquitous class of sulfated polysaccharides, playa critical role in various physiological processes, such as development,wound healing, angiogenesis, cell division, inflammation and spinal cordinjury. The complexity of this class of natural molecules has madecorrelating structure to function difficult. Synthetic glycopolymerswith tunable properties may offer an alternative to natural molecules.

SUMMARY OF THE INVENTION

In one aspect, the present disclosure provides a glycopolymer comprisinga plurality of repeating units, wherein each of the repeating unitscomprises a saccharide moiety (SA), a linking group (L) and a polymerbackbone moiety (PB). The repeating units can be optionally connected byone or more carbon-carbon double bonds. In some cases, the glycopolymercan be of the formula:

wherein:

n can be an integer between 1 and 1000000;

R′ and R″ can each be independently selected from the group consistingof unsubstituted or substituted alkyl, unsubstituted or substitutedheteroalkyl, unsubstituted or substituted aryl, unsubstituted orsubstituted heteroaryl, unsubstituted or substituted cycloalkyl, andunsubstituted or substituted heterocycloalkyl; and

the ratio of the total number of carbon-carbon double bonds in theglycopolymer to the total number of the repeating units can be less thanabout 1, with the proviso that the polymer backbone moiety may notcomprise a tetrahydrofuranyl.

In some cases, the glycopolymer can be of the formula:

In some cases, at least about 80% of the saccharide moiety in therepeating units comprises a heparan sulfate (HS) unit.

In another aspect, the present disclosure provides a glycopolymercomprising a plurality of repeating units, wherein each of the repeatingunits comprises a saccharide moiety (SA), a linking group (L) and apolymer backbone moiety (PB). The repeating units can be optionallyconnected by one or more carbon-carbon double bonds. In some cases, theglycopolymer is of the formula:

wherein:

n can be an integer between 1 and 1000000;

R′ and R″ can each be independently selected from the group consistingof unsubstituted or substituted alkyl, unsubstituted or substitutedheteroalkyl, unsubstituted or substituted aryl, unsubstituted orsubstituted heteroaryl, unsubstituted or substituted cycloalkyl, andunsubstituted or substituted heterocycloalkyl; and the saccharide moietycan comprises one or more heparan sulfate (HS) units.

In some cases, the saccharide moiety is of the formula:

wherein R₁, R₂, R₄, R₅ and R₆ are each independently selected from thegroup consisting of hydrogen, hydroxyl, sulfite, sulfate, acetyl,phosphate and carboxylate; and R₃ is hydroxyl. In some cases, R₂ can besulfite. In further cases, each of R₁ and R₄ can be independentlysulfate; and R₂ can be sulfite. In some cases, R₅ and R₆ can behydroxyl.

In yet another aspect, the present disclosure provides a glycopolymercomprising a plurality of repeating units, wherein each of the repeatingunits comprises a saccharide moiety (SA), a linking group (L) and apolymer backbone moiety (PB). The repeating units can be optionallyconnected by one or more carbon-carbon double bonds. In some cases, theglycopolymer can be of the formula:

wherein:

n can be an integer between 1 and 1000000;

R′ and R″ can each be independently selected from the group consistingof unsubstituted or substituted alkyl, unsubstituted or substitutedheteroalkyl, unsubstituted or substituted aryl, unsubstituted orsubstituted heteroaryl, unsubstituted or substituted cycloalkyl, andunsubstituted or substituted heterocycloalkyl; and the saccharide moietycan consist of three sulfate groups.

In various embodiments, the saccharide moiety can selected from thegroup consisting of a monosaccharide, a disaccharide, a trisaccharide, atetrasaccharide, a pentasaccharide and an oligosaccharide. For example,the saccharide moiety can comprise a disaccharide.

In some cases, the polydispersity of the glycopolymer can less thanabout 4. In some cases, the glycopolymer lacks anti-coagulant activity.In some cases, n can be greater than about 10.

In some cases, each of the repeating units can be of the formula:

wherein m is an integer between 1 and 1000. In some examples, m can be2.

In further cases, each of the repeating units can be of the formula:

wherein R₁, R₂ and R₄ are each independently selected from the groupconsisting of hydrogen, hydroxyl, sulfite, sulfate, acetyl, phosphateand carboxylate. In some examples, m can be 2. In some examples, R₁ andR₄ can each be sulfate; and R₂ can be sulfite.

In one aspect, the present disclosure provides a method of treating aninflammatory condition in a subject in need thereof. The method cancomprise administering to the subject a therapeutically effective amountof a glycopolymer of the present disclosure.

In some cases, the inflammatory condition can be an acute systemicinflammatory disease or a chronic inflammatory disease. In some cases,the glycopolymer can inhibit RANTES binding as ascertained by acompetitive enzyme-linked immunosorbent assay. In some examples, theglycopolymer can inhibit RANTES binding up to about 91% as ascertainedby the competitive enzyme-linked immunosorbent assay.

In another aspect, the present disclosure provides a compositioncomprising a substantially homogeneous population of a glycopolymer ofthe present disclosure. In some cases, the composition can comprisegreater than 50% (w/w) of the glycopolymer. In some cases, thecomposition can comprise greater than 80% (w/w) of the glycopolymer. Infurther cases, the composition can comprise greater than 95% (w/w) ofthe glycopolymer.

In yet another aspect, the present disclosure provides a pharmaceuticalcomposition comprising a glycopolymer of the present disclosure, and apharmaceutically acceptable carrier.

In one aspect, the present disclosure provides a pharmaceuticalcomposition comprising a glycopolymer of the present disclosure, and asecond pharmaceutical agent.

In another aspect, the present disclosure provides a kit comprising aglycopolymer of the present disclosure.

In yet another aspect, the present disclosure provides a substrate,wherein a glycopolymer of the present disclosure is immobilized thereon.In some cases, the substrate can comprise a solid support. In somecases, the substrate can be an array.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present invention will be obtained by reference to thefollowing detailed description that sets forth illustrative embodiments,in which the principles of the invention are utilized, and theaccompanying drawings of which:

FIG. 1a provides schematics of heparin/heparan sulfate (HS)polysaccharides and their synthetic mimetic.

FIG. 1b shows an exemplary route for the retrosynthesis ofglycopolymers.

FIG. 2 shows an exemplary method for synthesizing compounds.

FIG. 3 provides a schematic illustration of an exemplary method for thesynthesis of glycopolymers.

FIGS. 4a and 4b show the RANTES binding abilities and relative FXaactivity of disaccharide, heparin and glycopolymers.

FIGS. 5a and 5b show the RANTES-induced migration of CCR3- andCCR5-expressing cells.

FIG. 6 shows the RANTES-induced migration of CCR3-expressing cells.

FIG. 7 shows the RANTES binding to CCR3-expressing cells.

FIG. 8 depicts the chemical structure of Arixtra, Org31550, andsynthetic glycopolymers 26 and 27.

FIG. 9 shows an exemplary method for retrosynthesizing glycopolymer 26from a disaccharide.

FIG. 10 shows an exemplary method for synthesizing (A) apolymer-conjugated acceptor and (B) a glycosyl phosphate donor.

FIG. 11 provides an exemplary synthetic route for glycopolymer 26.

FIGS. 12A and 12B show the relative FXa and FIIa activity of heparin,LMWH and Arixtra.

FIGS. 13A and 13B show the relative FXa and FIIa activity of heparin andglycopolymers.

FIG. 14 shows the neutralization of the anti-FIIa activity of heparinand glycopolymers by PF4.

DETAILED DESCRIPTION

Glycosaminoglycans such as heparan sulfate (HS) participate in criticalbiological processes by modulating the activity of a diverse set ofprotein binding partners. Such proteins include all known members of thechemokine superfamily, which are thought to guide the migration ofimmune cells through their interactions with HS. A major challenge tounderstanding the structure—activity relationship of HS and developingHS-based therapeutic approaches has been the chemical complexity andheterogeneity of HS in vivo. Heparin, a close structural relative of HS,displays less heterogeneity and is used clinically as an anti-coagulantdrug for the prevention and treatment of thrombosis. Elegant studieshave demonstrated that a unique sulfated sequence found within heparinis primarily responsible for its anti-coagulant activity. Heparin hasalso been shown to have potent anti-inflammatory activity in models ofasthma, chronic dermatitis, and ulcerative colitis, but it is notrecommended as an anti-inflammatory agent in clinical practice due toits anti-coagulant activity.

Moreover, heparin isolated from natural sources can induce otherundesirable physiological effects due to its structural complexity andheterogeneity. First, it carries the potential risk of contamination.The global distribution of contaminated heparin in 2007, for example,resulted in over 100 deaths and hundreds of additional cases reportingadverse clinical effects. Second, heparin displays a variabledose-response relationship among patients, in part due to its structuralheterogeneity, and thus often requires active monitoring to fine-tunethe dosages. Lastly, approximately 3% of patients undergoing prolongedheparin therapy experience heparin-induced thrombocytopenia (HIT), asevere autoimmune response triggered by the complex formation of heparinand platelet factor 4 (PF4). Therefore, there is a need to develop saferalternatives to natural-sourced glycosaminoglycans that have morepredictable bioactivity and reduced side effects.

As used herein, the singular form “a”, “an”, and “the” include pluralreferences unless the context clearly dictates otherwise.

As used herein, the term “glycopolymer” refers to any polymer thatcomprises one or more saccharide moieties, for example, apolysaccharide, or a glycosaminoglycan.

As used herein, the term “effective amount” or “therapeuticallyeffective amount” refers to that amount of a compound described hereinthat is sufficient to effect the intended application including but notlimited to disease treatment, as defined below. The therapeuticallyeffective amount may vary depending upon the intended application (invitro or in vivo), or the subject and disease condition being treated,e.g., the weight and age of the subject, the severity of the diseasecondition, the manner of administration and the like, which can readilybe determined by one of ordinary skill in the art. The term also appliesto a dose that will induce a particular response in target cells, e.g.reduction of platelet adhesion and/or cell migration. The specific dosewill vary depending on the particular compounds chosen, the dosingregimen to be followed, whether it is administered in combination withother compounds, timing of administration, the tissue to which it isadministered, and the physical delivery system in which it is carried.

The term “treatment”, “treating”, “palliating” or “ameliorating” areused interchangeably herein and can refer to an approach for obtainingbeneficial or desired results including but not limited to therapeuticbenefit and/or a prophylactic benefit. Therapeutic benefit can meaneradication or amelioration of the underlying disorder being treated. Atherapeutic benefit can be achieved with the eradication or ameliorationof one or more of the physiological symptoms associated with theunderlying disorder such that an improvement is observed in the patient,notwithstanding that the patient may still be afflicted with theunderlying disorder. For prophylactic benefit, the compositions may beadministered to a patient at risk of developing a particular disease, orto a patient reporting one or more of the physiological symptoms of adisease, even though a diagnosis of this disease may not have been made.For purposes of this invention, beneficial or desired clinical resultsinclude, but are not limited to, one or more of the following: reducingthe proliferation of, or destroying, cancerous cells or other diseasedcells, reducing metastasis of cancerous cells found in cancers,shrinking the size of the tumor, decreasing symptoms resulting from thedisease, increasing the quality of life of those suffering from thedisease, palliating the pain resulting from the disease, decreasing thedose of other medications required to treat the disease, delaying theprogression of the disease, and/or prolonging survival of individuals.Treatment can include preventing the disease, that is, causing theclinical symptoms of the disease not to develop by administration of aprotective composition prior to the induction of the disease;suppressing the disease, that is, causing the clinical symptoms of thedisease not to develop by administration of a protective compositionafter the inductive event but prior to the clinical appearance orreappearance of the disease; inhibiting the disease, that is, arrestingthe development of clinical symptoms by administration of a protectivecomposition after their initial appearance; preventing re-occurring ofthe disease and/or relieving the disease, that is, causing theregression of clinical symptoms by administration of a protectivecomposition after their initial appearance.

As used herein, “agent” or “biologically active agent” refers to abiological, pharmaceutical, or chemical compound or other moiety.Non-limiting examples include simple or complex organic or inorganicmolecule, a peptide, a protein, an oligonucleotide, an antibody, anantibody derivative, antibody fragment, a vitamin derivative, acarbohydrate, a glycosaminoglycan, a glycomimetic, a toxin, or achemotherapeutic compound. Various compounds can be synthesized, forexample, small molecules and oligomers (e.g., oligopeptides andoligonucleotides), and synthetic organic compounds based on various corestructures. In addition, various natural sources can provide compoundsfor screening, such as plant or animal extracts, and the like. A skilledartisan can readily recognize that there is no limit as to thestructural nature of the agents of the present invention.

As used here, the term “pharmaceutically acceptable” refers to thosecompounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

As used here, the term “pharmaceutically-acceptable carrier” means apharmaceutically-acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, manufacturing aid (e.g.,lubricant, talc magnesium, calcium or zinc stearate, or steric acid), orsolvent encapsulating material, involved in carrying or transporting thecompound from one organ, or portion of the body, to another organ, orportion of the body. Each carrier must be “acceptable” in the sense ofbeing compatible with the other ingredients of the formulation and notinjurious to the subject.

As used herein, the term “pharmaceutically acceptable salts” is meant toinclude salts of active compounds which are prepared with relativelynontoxic acids or bases, depending on the particular substituentmoieties found on the compounds described herein. When compounds of thepresent invention contain relatively acidic functionalities, baseaddition salts can be obtained by contacting the neutral form of suchcompounds with a sufficient amount of the desired base, either neat orin a suitable inert solvent. Examples of pharmaceutically acceptablebase addition salts include sodium, potassium, calcium, ammonium,organic amino, or magnesium salt, or a similar salt. When compounds ofthe present invention contain relatively basic functionalities, acidaddition salts can be obtained by contacting the neutral form of suchcompounds with a sufficient amount of the desired acid, either neat orin a suitable inert solvent. Examples of pharmaceutically acceptableacid addition salts include those derived from inorganic acids likehydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic,phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, maleic, malonic, benzoic, succinic,suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic,p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Alsoincluded are salts of amino acids such as arginate and the like, andsalts of organic acids like glucuronic or galactunoric acids and thelike (see, for example, Berge et al., “Pharmaceutical Salts”, Journal ofPharmaceutical Science, 1977, 66, 1-19). Certain specific compounds ofthe present invention contain both basic and acidic functionalities thatallow the compounds to be converted into either base or acid additionsalts.

As used herein, the term “homogeneous” refers to a mixture or blend ofcomponents that is generally uniform in structure and composition withlittle variability throughout the mixture. Different portions of ahomogeneous mixture exhibit the same physical and chemical properties atevery place throughout the mixture. The stoichiometry in a homogeneousmixture is also constant throughout the mixture.

As used herein, the term “anti-inflammatory activity” means an abilityto reduce or prevent one or more biological processes associated withinflammatory events.

As used herein, the term “anti-coagulant activity” we mean an ability toreduce or prevent coagulation (i.e. the clotting of blood) or anassociated signal or effect.

As used herein, the term “administer” or “administering” refers to theplacement of a composition into a subject by a method or route whichresults in at least partial localization of the composition at a desiredsite such that desired effect is produced. A compound or compositiondescribed herein can be administered by any appropriate route known inthe art including, but not limited to, oral or parenteral routes,including intravenous, intramuscular, subcutaneous, transdermal, airway(aerosol), pulmonary, nasal, rectal, and topical (including buccal andsublingual) administration.

When ranges are used herein for physical properties, such as molecularweight, or chemical properties, such as chemical formulae, allcombinations and subcombinations of ranges and specific embodimentstherein are intended to be included. The term “about” when referring toa number or a numerical range means that the number or numerical rangereferred to is an approximation within experimental variability (orwithin statistical experimental error), and thus the number or numericalrange may vary from, for example, between 1% and 15% of the statednumber or numerical range. The term “comprising” (and related terms suchas “comprise” or “comprises” or “having” or “including”) includes thoseembodiments, for example, an embodiment of any composition of matter,composition, method, or process, or the like, that “consist of” or“consist essentially of” the described features.

As used herein, a “subject” means a human or an animal, such as apatient, a domesticated animal (e.g., dog, cat, and the like), a farmanimal (e.g., cow, sheep, pig, horse, and the like) or a laboratoryanimal (e.g., rat, mouse, guinea pig, and the like).

As used herein, the term “in vivo” refers to an event that takes placein a subject's body.

As used herein, the term “in vitro” refers to an event that takes placesoutside of a subject's body. For example, an in vitro assay mayencompass any assay run outside of a subject assay. In vitro assays mayencompass cell-based assays in which cells alive or dead are employed.In vitro assays may also encompass a cell-free assay in which no intactcells are employed.

Description of compounds of the present invention is limited byprinciples of chemical bonding known to those skilled in the art.Accordingly, where a group may be substituted by one or more of a numberof substituents, such substitutions are selected so as to comply withprinciples of chemical bonding and to give compounds which are notinherently unstable and/or would be known to one of ordinary skill inthe art as likely to be unstable under ambient conditions, such asaqueous, neutral, and several known physiological conditions. Forexample, a heterocycloalkyl or heteroaryl is attached to the remainderof the molecule via a ring heteroatom in compliance with principles ofchemical bonding known to those skilled in the art thereby avoidinginherently unstable compounds.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds which differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement of a hydrogen by a deuterium ortritium, or the replacement of a carbon by 13C- or 14C-enriched carbonare within the scope of this invention.

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of atoms that constitutesuch compounds. For example, the compounds may be radiolabeled withradioactive isotopes, such as for example tritium (3H), iodine-125(1251) or carbon-14 (14C). All isotopic variations of the compounds ofthe present invention, whether radioactive or not, are encompassedwithin the scope of the present invention.

As used herein, the term “alkyl” refers to a straight or branchedhydrocarbon chain radical consisting solely of carbon and hydrogenatoms, containing no unsaturation, having from one to one hundred carbonatoms (e.g., C1-C100 alkyl). Whenever it appears herein, a numericalrange such as “1 to 100” refers to each integer in the given range;e.g., “1 to 100 carbon atoms” means that the alkyl group may consist of1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including100 carbon atoms, although the present definition also covers theoccurrence of the term “alkyl” where no numerical range is designated.Typical alkyl groups include, but are in no way limited to, methyl,ethyl, propyl, isopropyl, n-butyl, iso-butyl, sec-butyl isobutyl,tertiary butyl, pentyl, isopentyl, neopentyl, hexyl, septyl, octyl,nonyl, decyl, and the like. The alkyl is attached to the rest of themolecule by a single bond, for example, methyl (Me), ethyl (Et),n-propyl, 1-methylethyl (iso propyl), n-butyl, n-pentyl,1,1-dimethylethyl (t butyl), 3-methylhexyl, 2-methylhexyl, and the like.An alkyl moiety may be unsubstituted or substituted.

As used herein, the term “heteroalkyl” refers to a straight- orbranched-chain alkyl group preferably having from 2 to 100 carbons, oneor more of which has been replaced by a heteroatom selected from S, O, Pand N. Exemplary heteroalkyls include alkyl ethers, secondary andtertiary alkyl amines, amides, alkyl sulfides, and the like. The groupmay be a terminal group or a bridging group. A heteroalkyl moiety may beunsubstituted or substituted.

As used herein, the term “aromatic” or “aryl” refers to an aromaticradical with three to sixteen carbon atoms (e.g., —C3-16aromatic or—C3-16aryl) which has at least one ring having a conjugated pi electronsystem which is carbocyclic (e.g., phenyl, fluorenyl, and naphthyl).Whenever it appears herein, a numerical range such as “C3-10” refers toeach integer in the given range; e.g., “—C3-10aryl” means that the arylgroup may consist of 3 ring atoms, 4 ring atoms, etc., up to andincluding 10 ring atoms. The term includes monocyclic or fused-ringpolycyclic (i.e., rings which share adjacent pairs of ring atoms)groups. Examples of aryl include, but are not limited to, phenyl,4-chlorophenyl, 4-fluorophenyl, 4-bromophenyl, 3-nitrophenyl,2-methoxyphenyl, 2-methylphenyl, 3-methyphenyl, 4-methylphenyl,4-ethylphenyl, 2-methyl-3-methoxyphenyl, 2,4-dibromophenyl,3,5-difluorophenyl, 3,5-dimethylphenyl, 2,4,6-trichlorophenyl,4-methoxyphenyl, naphthyl, 2-chloronaphthyl, 2,4-dimethoxynaphthyl,4-(trifluoromethyl)naphthyl, 2-iodo-4-methylnaphthyl, and the like. Anaryl moiety may be unsubstituted or substituted.

As used herein, the term “heteroaryl” or, alternatively,“heteroaromatic”, “hetaryl”, ‘heteroar” or “hetar” refers to an aromaticradical with one to sixteen carbon atoms (e.g., —C1-16heteroaryl) thatfurther includes one or more ring heteroatoms selected from nitrogen,oxygen and sulfur, and which may be a monocyclic, bicyclic, tricyclic ortetracyclic ring system. Whenever it appears herein, a numerical rangesuch as “C1-10” refers to each integer in the given range; e.g.,“—C1-10hetaryl” means that the heteroaryl group may consist of 1 carbonatoms, 2 carbon atoms, etc., up to and including 10 carbon atoms. An“N-containing heteroaromatic” or “N-containing heteroaryl” moiety refersto an aromatic group in which at least one of the skeletal atoms of thering is a nitrogen atom. The polycyclic heteroaryl group may be fused ornon-fused. The heteroatom(s) in the heteroaryl radical is optionallyoxidized. One or more nitrogen atoms, if present, are optionallyquaternized. The heteroaryl is attached to the rest of the moleculethrough any atom of the ring(s). A heteroaryl moiety may beunsubstituted or substituted.

As used herein, the term “cycloalkyl” refers to a saturated or partiallyunsaturated ring structure with three to ten carbon atoms (i.e.—C3-10cycloalkyl). Whenever it appears herein, a numerical range such as“C3-10” refers to each integer in the given range; e.g.,“—C3-10cycloalkyl” means that the cycloalkyl group may consist of 3carbon atoms, 4 carbon atoms, etc., up to and including 10 carbon atoms.The term includes monocyclic or fused-ring polycyclic (i.e., rings whichshare adjacent pairs of ring atoms) groups. A cycloalkyl moiety may beunsubstituted or substituted.

As used herein, the term “heterocyclyl”, “hetcyclyl”, or“heterocycloalkyl” refers to a 3-, 4-, 5-, or 6-membered saturated orpartially unsaturated ring containing one, two, or three heteroatoms,preferably one or two heteroatoms independently selected from oxygen,nitrogen and sulfur; or to a bicyclic ring system containing up to 10atoms including at least one heteroatom independently selected fromoxygen, nitrogen, and sulfur wherein the ring containing the heteroatomis saturated. Whenever it appears herein, a numerical range such as“C1-10” refers to each integer in the given range; e.g.,“—C1-10heterocyclyl” means that the heterocycloalkyl group may consistof 1 carbon atoms, 2 carbon atoms, etc., up to and including 10 carbonatoms. Examples of heterocyclyl include, but are not limited to,tetrahydrofuranyl, tetrahydrofuryl, pyrrolidinyl, piperidinyl,4-pyranyl, tetrahydropyranyl, thiolanyl, morpholinyl, piperazinyl,dioxolanyl, dioxanyl, indolinyl, and chromanyl. A hetereocycloalkylmoiety may be unsubstituted or substituted.

As used herein, the term “alkoxy” denotes an optionally substitutedstraight or branched chain alkyl-oxy group wherein the “alkyl” portionis as defined such as methoxy, ethoxy, n-propyloxy, i-propyloxy,n-butyloxy, i-butyloxy, tert.-butyloxy, pentyloxy, hexyloxy, heptyloxyincluding their isomers.

As used herein, the term “substituted” means that the referenced groupmay be substituted with any one or more additional chemical group(s)known in the art. Examples of such chemical groups include, but are notlimited to, alkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, hydroxy,alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, halo, amino(unsubstituted, or mono- or disubstituted), acyl, carbonyl, carboxyl,ester, amido, thiocarbonyl, isocyano, thiocyano, isothiocyano, nitro,perhaloalkyl, (e.g. perfluoroalkyl), phosphate, silyl, sulfinyl,sulfonyl, sulfonamidyl, sulfoxyl, sulfonate, carbohydrates, and anyprotected derivatives thereof. The substituents themselves may besubstituted, for example, a cycloalkyl substituent may have a halidesubstituted at one or more ring carbons, and the like. The protectinggroups that may form the protected derivatives of the above substituentsare known to those of skill in the art and may be found in referencessuch as Wuts P.; Greene T., (Greene's Protective Groups in OrganicSynthesis, 4th Edition, 2006) which is herein incorporated by referencein its entirety.

Compounds described herein can contain one or more asymmetric centersand may thus give rise to diastereomers and optical isomers. The presentdisclosure includes all such possible diastereomers as well as theirracemic mixtures, their substantially pure resolved enantiomers, allpossible geometric isomers, and pharmaceutically acceptable saltsthereof. Furthermore, mixtures of stereoisomers as well as isolatedspecific stereoisomers are also included. During the course of thesynthetic procedures used to prepare such compounds, or in usingracemization or epimerization procedures known to those skilled in theart, the products of such procedures can be a mixture of stereoisomers.

The present disclosure includes all manner of rotamers andconformationally restricted states of a compound of the invention.

Compounds

I. General Structure

In one aspect, the present disclosure provides a glycopolymer. Theglycopolymer can comprise a plurality of repeating units, wherein eachof the repeating units comprises a saccharide moiety (SA), a linkinggroup (L) and a polymer backbone moiety (PB). The repeating units can beoptionally connected by one or more carbon-carbon double bonds. Forexample, the glycopolymer can be of the formula:

wherein n is an integer between 1 and 1000000; R′ and R″ are eachindependently selected from the group consisting of unsubstituted orsubstituted alkyl, unsubstituted or substituted heteroalkyl,unsubstituted or substituted aryl, unsubstituted or substitutedheteroaryl, unsubstituted or substituted cycloalkyl, and unsubstitutedor substituted heterocycloalkyl.

In some cases, n can be an integer greater than 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 60,70, 80, 90, 100, 150, 200, 250, 300, 400, 500, 600, 700, 800, 900, 1000,1500, 2000, 2500, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 10000,15000, 20000, 25000, 30000, 40000, 50000, 60000, 70000, 80000, 90000,100000, 150000, 200000, 250000, 300000, 400000, 500000, 600000, 700000,800000, 900000, or 1000000. For example, n may be greater than 10 or 45.Alternatively, n can be an integer less than 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 60, 70,80, 90, 100, 150, 200, 250, 300, 400, 500, 600, 700, 800, 900, 1000,1500, 2000, 2500, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 10000,15000, 20000, 25000, 30000, 40000, 50000, 60000, 70000, 80000, 90000,100000, 150000, 200000, 250000, 300000, 400000, 500000, 600000, 700000,800000, 900000, or 1000000. For example, n can be smaller than 1000 or500. In some cases, n may be in a range between any of the two valuesdescribed herein. For example, n can be 48 or 155.

In some cases, R′ and R″ can be each independently selected from thegroup consisting of unsubstituted or substituted alkyl, unsubstituted orsubstituted aryl, and unsubstituted or substituted cycloalkyl. In othercases, R′ and R″ are each independently selected from the groupconsisting of unsubstituted or substituted heteroalkyl, unsubstituted orsubstituted heteroaryl, and unsubstituted or substitutedheterocycloalkyl. In some cases, R′ and R″ are each independentlyselected from the group consisting of unsubstituted or substitutedalkyl, unsubstituted or substituted heteroalkyl, unsubstituted orsubstituted aryl, and unsubstituted or substituted heteroaryl. In someexamples, R′ and R″ can independently be unsubstituted or substitutedalkyl. For example, R′ can be methyl and R″ can be benzyl (i.e. a methylsubstituted with a phenyl). In some examples, R′ can be unsubstituted orsubstituted aryl, and R″ can be unsubstituted or substitutedheteroalkyl. For example, R′ can be 3-chloro-4-nitro-phenyl and R″ canbe a polyethylene glycol (PEG) linker substituted with another polymer(e.g. a glycopolymer). In some cases, the glycopolymer can be linked toother polymer(s) via the R′ and/or R″ to form a block copolymer.

In some cases, the repeating units can each be connected to twocarbon-carbon double bonds. In some cases, the carbon-carbon doublebonds can be fully or partially reduced. The ratio of the total numberof carbon-carbon double bonds in the glycopolymer to the total number ofthe repeating units can be less than about 1. The total number ofcarbon-carbon double bonds can be less than the total number ofrepeating units. In further cases, the carbon-carbon double bonds can befully reduced. The repeating units can each be connected to zerocarbon-carbon double bonds. In some cases, the polymer backbone moietymay not comprise a tetrahydrofuranyl group, such as in the Ring-OpeningMetathesis Polymerization (ROMP) product of an oxanorbornene monomer

In some cases, the saccharide moiety may comprise one or more heparansulfate (HS) units, which are known to one skilled in the art. Heparansulfate (HS) units can comprise a first unit [e.g. β-D-glucuronic acid(GlcA) or α-L-iduronic acid (IdoA)] linked to a second unit [e.g.2-deoxy-2-acetamido-α-D-glucopyranosy (GlcNAc) or2-deoxy-2-sulfamido-α-D-glucopyranosyl (GlcNS)], wherein the first unitand/or second unit can be optionally substituted with one, two, three ormore sulfate groups. For example, a heparan sulfate unit can be of theformula:

wherein R₁, R₂, R₄, R₅, R₆ may each be independently hydrogen, hydroxyl,sulfite, sulfate, phosphate, acetyl or carboxylate.

The saccharide moiety may comprise three or more sulfate groups. Forexample, the saccharide moiety may consist of three sulfate groups.Alternatively, the saccharide moiety can comprise four, five, six, ormore sulfate groups.

II. Polymer Backbone Moiety (PB)

In some cases, the polymer backbone moiety can be derived from astrained ring system that can be used for Ring-Opening MetathesisPolymerization (ROMP). Examples include but are not limited tocyclobutene, cyclopentene, cyclooctene, cyclooctatetraene, norbornene,oxonorbornene, dicyclopentadiene, or substituted derivatives thereof.

In some cases, the polymer backbone moiety may be a natural polymermoiety. As used herein, “natural polymer” refers to polymers that existin nature, such polymers found in plants, animals and humans. In somecases, the polymer backbone moiety may be a synthetic polymer moiety. Asused herein, “synthetic polymer” refers to polymers that areartificially produced by chemical processes. In some cases, the polymerbackbone moiety may be a combination of natural polymer moieties andsynthetic polymer moieties. Examples of natural polymer moieties includebut are not limited to amino acids, saccharides (e.g. monosaccharides,dissacharides), nucleic acids, and other natural polymers, orcombinations thereof. Non-limiting examples of synthetic polymermoieties may include moieties of acrylate, nylon, silicone, spandex,viscose rayon, carboxylic acid, vinyl acetate, acrylamide, ethyleneglycol, cyclobutene, cyclopentene, norbornene, cyclooctene,cyclooctatetraene, urethane, lactic acid, silica, styrene,acrylonitrile, butadiene, carbonate, ethylene, ethylene terephthalate,chlorotrifluoroethylene, ethylene oxide, ethylene terephthalate,ethylene, isobutylene, methyl methacrylate, oxymethylene, formaldehyde,propylene, tetrafluoroethylene, vinyl acetate, vinyl alcohol, vinylchloride, vinylidene dichloride, vinylidene difluoride, vinyl fluoride,or combinations (e.g., block copolymers, alternating copolymers, randomcopolymers etc.) thereof.

In some cases, the polymer backbone moiety can comprise a cyclopentylgroup. For example, the glycopolymer may be of the formula:

wherein R′ and R″ are each independently selected from the groupconsisting of unsubstituted or substituted alkyl, unsubstituted orsubstituted heteroalkyl, unsubstituted or substituted aryl,unsubstituted or substituted heteroaryl, unsubstituted or substitutedcycloalkyl, and unsubstituted or substituted heterocycloalkyl.

In some cases, the polymer backbone moiety may not comprise atetrahydrofuranyl group, such as in the Ring-Opening MetathesisPolymerization (ROMP) product of 7-oxonorbornene.

Polymer backbone moiety may be biodegradable. By “biodegradable” we meanthe absorbability or degradation of a compound or composition whenadministered in vivo or under in vitro conditions. Biodegradation mayoccur through the action of biological agents, either directly orindirectly. Examples of biodegradable polymer backbone moieties includebut are not limited to lactides, glycolide, trimethylene carbonate,lactide-co-glycolide, ethylene terephtalate, caprolactone, catgut suturematerial, collagen (e.g., equine collagen foam), lactic acid, orhyaluronic acid.

In some cases, a certain percentage of the overall population of therepeating units may comprise natural polymer backbone moieties. Forexample, in some cases, none of the repeating units may comprise naturalpolymer backbone moieties. In some cases, about 1%, 2%, 3%, 4%, 5%, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%,85%, 90%, 95%, or 100% of the repeating units may comprise naturalpolymer backbone moieties. In one example, about 10% of the repeatingunits may comprise natural polymer backbone moieties. In anotherexample, about 50% of the repeating units may comprise natural polymerbackbone moieties. In some cases, more than about 1%, 2%, 3%, 4%, 5%,10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%,80%, 85%, 90%, 95%, or 99% of the repeating units may comprise naturalpolymer backbone moieties. For example, more than about 50% or 75% ofthe repeating units may comprise natural polymer backbone moieties. Insome cases, less than about 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%,35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%of the repeating units may comprise natural polymer backbone moieties.For example, less than about 25% or 50% of the repeating units maycomprise natural polymer backbone moieties. In some cases, thepercentage of repeating units that comprise natural polymer backbonemoieties may fall between any of the two values described herein. Forexample, about 99% of the repeating units may comprise natural polymerbackbone moieties.

In some cases, a certain percentage of the overall population of therepeating units may comprise synthetic polymer backbone moieties. Forexample, in some cases, none of the repeating units may comprisesynthetic polymer backbone moieties. In some cases, about 1%, 2%, 3%,4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,75%, 80%, 85%, 90%, 95%, or 100% of the repeating units may comprisesynthetic polymer backbone moieties. In one example, about 100% of therepeating units may comprise synthetic polymer backbone moieties. Inanother example, about 85% of the repeating units may comprise syntheticpolymer backbone moieties. In some cases, at least about 1%, 2%, 3%, 4%,5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,75%, 80%, 85%, 90%, 95%, or 99% of the repeating units may comprisesynthetic polymer backbone moieties. For example, at least about 75% or90% of the repeating units may comprise synthetic polymer backbonemoieties. In some cases, no more than about 1%, 2%, 3%, 4%, 5%, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%,85%, 90%, 95%, or 100% of the repeating units may comprise syntheticpolymer backbone moieties. For example, less than about 25% or 10% ofthe repeating units may comprise synthetic polymer backbone moieties. Insome cases, the percentage of repeating units that comprise syntheticpolymer backbone moieties may fall between any of the two valuesdescribed herein. For example, about 99% of the repeating units maycomprise synthetic polymer backbone moieties.

In some cases, it may be desirable to have glycopolymers with apopulation of repeating units that comprise identical polymer backbonemoieties. For example, in some cases, about 1%, 2%, 3%, 4%, 5%, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%,85%, 90%, 95%, or 100% of the repeating units may comprise identicalpolymer backbone moieties. In one example, about 50% of the repeatingunits may comprise identical polymer backbone moieties. In anotherexample, about 100% of the repeating units may comprise polymer backbonemoieties. In some cases, at least about 1%, 2%, 3%, 4%, 5%, 10%, 15%,20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95%, or 99% of the repeating units may comprise identical polymerbackbone moieties. For example, at least about 75% or 90% of therepeating units may comprise identical polymer backbone moieties. Insome cases, no more than about 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%,30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or100% of the repeating units may comprise identical polymer backbonemoieties. For example, no more than 10% or 50% of the repeating unitsmay comprise identical polymer backbone moieties. In some cases, thepercentage of repeating units that comprise identical polymer backbonemoieties may fall between any of the two values described herein. Forexample, about 99.5% or 99.9% of the repeating units may compriseidentical polymer backbone moieties.

Additionally or alternatively, in some cases, glycopolymers with apopulation of repeating units that comprise differing polymer backbonemoieties may be prepared. In some cases, about 1%, 2%, 3%, 4%, 5%, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%,85%, 90%, 95%, or 100% of the repeating units may comprise differingpolymer backbone moieties. For example, about 5% or 50% of the repeatingunits may comprise differing polymer backbone moieties. In some cases,at least about 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of therepeating units may comprise differing polymer backbone moieties. Forexample, at least about 1% or 10% of the repeating units may comprisediffering polymer backbone moieties. In some cases, no more than about1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the repeating unitsmay comprise differing polymer backbone moieties. For example, no morethan 25% or 50% of the repeating units may comprise differing polymerbackbone moieties. In some cases, the percentage of repeating units thatcomprise differing polymer backbone moieties may fall between any of thetwo values described herein. For example, about 0.5% of the repeatingunits may comprise differing polymer backbone moieties.

The polymer backbone moiety may be fully saturated, partially saturated,or unsaturated. A “partially saturated” polymer backbone moietycomprises, or is directly connected to, at least one carbon-carbon bondis unsaturated (e.g., a carbon-carbon double bond, a carbon-carbontriple bond). To modify the properties (e.g., conformation, rigidity,flexibility) of the glycopolymer, one or more of the carbon-carbondouble bonds may be reduced. The degree of reduction may be determinedby the ratio between total number of carbon-carbon double bonds inpolymer backbone moieties and the number of repeating units. Forexample, if all of the carbon-carbon double bonds are reduced, the ratiois zero. Alternatively, if each of the polymer backbone moieties isconnected to one or more carbon-carbon double bonds, then the ratio isgreater than 1.

Based on the desired properties of glycopolymers, this ratio may bevaried. In some cases, the ratio may be zero. In some cases, the ratiomay be about 0.000001, 0.000005, 0.00001, 0.00005, 0.0001, 0.0005,0.001, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1,0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1. For example, the ratio maybe about 0.0001 or 0.001. In some cases, the ratio may be more thanabout 0.000001, 0.000005, 0.00001, 0.00005, 0.0001, 0.0005, 0.001,0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2,0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1. For example, the ratio may bemore than about 0.0001 or 0.1. In some cases, the ratio may be less thanabout 0.000001, 0.000005, 0.00001, 0.00005, 0.0001, 0.0005, 0.001,0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2,0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1. For example, the ratio may beless than about 1 or 0.5. In some cases, the ratio may fall into a rangeof any of the two values described herein. For example, the ratio may be0.015 or 0.12.

To tune the properties of glycopolymers, the polymer backbone moiety maybe modified or functionalized (e.g., substituted, hydrothiolated,reduced, hydrogenated, hydrolyzed etc.). The polymer backbone moiety maybe modified or functionalized during the formation of the glycopolymer(i.e., simultaneously), or after the formation of the glycopolymer(i.e., sequentially).

III. Linking Group (L)

As described herein, a linking group may be any groups or bonds, throughwhich, the saccharide moiety are attached to the polymer backbonemoiety. The linking group may be selected from the groups consistingalkyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl,alkenylalkyl, alkynyl, alkynylalkyl, hydroxyalkyl, haloalkyl andheterocycloalkyl, which may or may not be substituted by one or more ofsubstituents. Examples of substituents may include, but not limited toalkyl, aryl, heterocyclyl, cycloalkyl, nitro, cyano, azido, amino, alkylamino, dialkyl amino, cycloalkyl amino, aryl amino, diarylamino,heterocyclyl amino, hydroxy, alkoxy, aryloxy, heterocyclyloxy,cycloalkoxy, thio, alkylthio, arylthio, heterocyclylthio, alkylcarbonyl, cycloalkyl carbonyl, aryl carbonyl, heterocyclyl carbonyl,carboxy, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,arylaminocarbonyl, dialkylaminocarbonyl, diarylaminocarbonyl,heterocyclylaminocarbonyl. In some cases, substituents may be furthersubstituted. In some cases, the linking group may be a bond (e.g. acarbon-carbon bond).

In some embodiments, the linking group may comprise a bifunctionallinker, a trifunctional linker, a multifunctional linker, or acombination thereof. Examples of linking groups may include but are notlimited to ethylene, ethylene glycol, oxyethylene, methylene glycol,trimethylene glycol, vinylpyrrolidones, Alkyne-PEG5-acid,N-Alloc-1,4-butandiamine hydrochloride, N-Alloc-1,6-hexanediaminehydrochloride, N-Alloc-1,3-propanediamine hydrochloride,4-Acetyl-3,5-dioxo-1-methylcyclohexanecarboxylic acid,Allyl(4-methoxyphenyl)dimethylsilane,6-(Allyloxycarbonylamino)-1-hexanol,3-(Allyloxycarbonylamino)-1-propanol, 4-Aminobutyraldehyde diethylacetal,(E)-N-(2-Aminoethyl)-4-{2-[4-(3-azidopropoxyl)phenyl]diazenyl}benzamidehydrochloride, N-(2-Aminoethyl)maleimide trifluoroacetate salt,Amino-PEG4-alkyne, Benzyl N-(3-hydroxypropyl)carbamate,4-(Boc-amino)-1-butanol, 4-(Boc-amino)butyl bromide,2-(Boc-amino)ethanethiol, 2-[2-(Boc-amino)ethoxy]ethoxyacetic acid(dicyclohexylammonium) salt, 2-(Boc-amino)ethyl bromide,6-(Boc-amino)-1-hexanol,21-(Boc-amino)-4,7,10,13,16,19-hexaoxaheneicosanoic acid purum,6-(Boc-amino)hexyl bromide, 5-(Boc-amino)-1-pentanol,3-(Boc-amino)-1-propanol, 3-(Boc-amino)-1-propanolpurum,3-(Boc-amino)propyl bromide,15-(Boc-amino)-4,7,10,13-tetraoxapentadecanoic acid purum,N-Boc-1,4-butanediamine, N-Boc-cadaverine, N-Boc-ethanolamine,N-Boc-ethylenediamine, N-Boc-2,2′-(ethylenedioxy)diethylamine,N-Boc-1,6-hexanediamine, N-Boc-1,6-hexanediamine hydrochloride purum,N-Boc-1,6-hexanediamine hydrochloride, N-Boc-4-isothiocyanatoaniline,N-Boc-4-isothiocyanatobutylamine, N-Boc-2-isothiocyanatoethylamine,N-Boc-3-isothiocyanatopropylamine, N-Boc-N-methylethylenediamine,N-Boc-m-phenylenediamine, N-Boc-p-phenylenediamine,2-(4-Boc-1-piperazinyl)acetic acid, N-Boc-1,3-propanediamine,N-Boc-1,3-propanediamine,N-Boc-N′-succinyl-4,7,10-trioxa-1,13-tridecanediamine,N-Boc-4,7,10-trioxa-1,13-tridecanediamine, N-(4-Bromobutyl)phthalimide,4-Bromobutyric acid, 4-Bromobutyryl chloride purum, 4-Bromobutyrylchloride, N-(2-Bromoethyl)phthalimide, N-(2-Bromoethyl)phthalimide,6-Bromo-1-hexanol, 6-Bromo-1-hexanol purum, 3-(Bromomethyl)benzoic acidN-succinimidylester, 4-(Bromomethyl)phenyl isothiocyanate,8-Bromooctanoic acid, 8-Bromo-1-octanol,4-(2-Bromopropionyl)phenoxyacetic acid, N-(3-Bromopropyl)phthalimide,4-(tert-Butoxymethyl)benzoic acid, tert-Butyl2-(4-{[4-(3-azidopropoxyl)phenyl]azo}benzamido)ethylcarbamate,tert-Butyl trans-17-bromo-4,7,10,13-tetraoxa-15-heptadecenoate,2-[2-(tert-Butyldimethylsilyloxy)ethoxy]ethanamine, tert-Butyl4-hydroxybutyrate, 4-(2-Chloropropionyl)phenylacetic acid,1,11-Diamino-3,6,9-trioxaundecane, di-Boc-cystamine, Diethylene glycolmonoallyl ether, 3,4-Dihydro-2H-pyran-2-methanol,4-[(2,4-Dimethoxyphenyl)(Fmoc-amino)methyl]phenoxyacetic acid,4-(Diphenylhydroxymethyl)benzoic acid, 4-(Fmoc-amino)-1-butanol,2-(Fmoc-amino)ethanol, 2-[2-(Fmoc-amino)ethoxy]ethylamine hydrochloride,2-(Fmoc-amino)ethyl bromide, 6-(Fmoc-amino)-1-hexanol,5-(Fmoc-amino)-1-pentanol, 3-(Fmoc-amino)-1-propanol,3-(Fmoc-amino)propyl bromide, N-Fmoc-2-bromoethylamine,N-Fmoc-1,4-butanediamine hydrobromide, N-Fmoc-cadaverine hydrobromide,N-Fmoc-ethylenediamine hydrobromide, N-Fmoc-1,6-hexanediaminehydrobromide, N-Fmoc-1,3-propanediamine hydrobromide,N-Fmoc-N″-succinyl-4,7,10-trioxa-1,13-tridecanediamine,(3-Formyl-1-indolyl)acetic acid, 6-Guanidinohexanoic acid,4-Hydroxybenzyl alcohol purum, 4-Hydroxybenzyl alcohol,N-(4-Hydroxybutyl)trifluoroacetamide,4′-Hydroxy-2,4-dimethoxybenzophenone, N-(2-Hydroxyethyl)maleimide,4-[4-(1-Hydroxyethyl)-2-methoxy-5-nitrophenoxy]butyric acid,N-(2-Hydroxyethyl)trifluoroacetamide,N-(6-Hydroxyhexyl)trifluoroacetamide, 4-Hydroxy-2-methoxybenzaldehyde,4-Hydroxy-3-methoxybenzyl alcohol, 4-(Hydroxymethyl)benzoic acid,4-Hydroxymethyl-3-methoxyphenoxyacetic acid,4-(4-Hydroxymethyl-3-methoxyphenoxy)butyric acid,4-(Hydroxymethyl)phenoxyacetic acid,3-(4-Hydroxymethylphenoxyl)propionic acid,N-(5-Hydroxypentyl)trifluoroacetamide, 4-(4′-Hydroxyphenylazo)benzoicacid, N-(3-Hydroxypropyl)trifluoroacetamide, 2-Maleimidoethyl mesylate,4-Mercapto-1-butanol, 6-Mercapto-1-hexanol, Phenacyl4-(bromomethyl)phenylacetate, 4-Sulfamoylbenzoic acid,N-Trityl-1,2-ethanediamine hydrobromide, 4-(Z-Amino)-1-butanol,6-(Z-Amino)-1-hexanol, 5-(Z-Amino)-1-pentanol, N—Z-1,4-Butanediaminehydrochloride, N—Z-Ethanolamine, N—Z-Ethylenediamine hydrochloridepurum, N—Z-Ethylenediamine hydrochloride, N—Z-1,6-hexanediaminehydrochloride, N—Z-1,5-pentanediamine hydrochloride,N—Z-1,3-Propanediamine hydrochloride, N¹,N⁴-Bis-Boc-spermidine,N¹,N⁵-Bis-Boc-spermidine, N-Boc-diethanolamine,N¹-Boc-2,2′-iminodiethylamine, N-Boc-iminodipropionic acid,N1-Boc-3,3′-iminodipropylamine, N,N″-Di-Z-diethylenetriamine, andderivatives thereof.

In some cases, the linking group may have one of the formulas selectedfrom: —O—(CH₂)_(m)—, —O—(CH₂)_(m)—X—, —NH—(CH₂)_(m)—, —NH—(CH₂)_(m)—X—,—O—(CH₂—CH₂—O)_(m)—, —O—(CH₂—CH₂—O)_(m)—X—, —NH—(CH₂—CH₂—O)_(m)—,—NH—(CH₂—CH₂—O)_(m)—X—, where X is O or S and m is between 1 and 1000.

The linking groups may be identical or varied. In some cases, all of therepeating units may comprise an identical linking group. In some cases,equal to or less than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%,99.9%, or 100% of the repeating units may comprise an identical linkinggroup. For example, equal to or less than about 75% or 95% of therepeating units may comprise identical linking groups. In some cases,more than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%,55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or100% of the repeating units may comprise an identical linking group. Forexample, more than about 50% or 90% of the repeating units may compriseidentical linking groups. In some cases, the repeating units comprisingan identical linking group may constitute a percentage of the wholepopulation falling between any of the two values described herein. Forexample, about 98.5% of the repeating units may comprise identicallinking groups.

Alternatively, in some cases, it may be advantageous to have repeatingunits that comprise differing linking groups. For example, in somecases, equal to or less than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%,40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%,86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,99.5%, 99.9%, or 100% of the repeating units may comprise differentlinking groups. For example, equal to or less than about 50% or 25% ofthe repeating units may comprise different linking groups. In somecases, more than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%,99.9%, or 100% of the repeating units may comprise different linkinggroups. For example, more than about 5% or 25% of the repeating unitsmay comprise different linking groups. In some cases, the percentage ofrepeating units that comprise different linking groups may be betweenany of the two values described herein. For example, about 2.5% or 12.5%of the repeating units may comprise different linking groups.

IV. Saccharide Moiety (SA)

As provided herein, the saccharide moiety may refer to any sugar orcarbohydrate. The saccharide moiety may be natural (e.g., heparin) orsynthetic. The saccharide moiety may be straight chain or cyclic, i.e.,mono-, di- and poly-, straight chain and cyclic saccharides. Thesaccharide moiety may be a monosaccharide, a disaccharide, atrisaccharide, a tetrasaccharide, a pentasaccharide, an oligosacchride(i.e., a saccharide moiety with about 6-40 sugar residues), apolysaccharide (i.e., a saccharide moiety with about 40-3000 sugarresidues), or a combination thereof. Non-limiting examples of saccharidemoieties may include glucose, fructose, galactose, sucrose, lactulose,lactose, maltose, trehalose, cellobiose, chondrotin sulfate, dermatansulfate, keratan sulfate, heparin, heparan sulfate, or combinationsthereof.

In some cases, the saccharide moieties may vary in the type of uronicsugar (e.g., glucuronic acid, iduronic acid, galactose etc.) and aminosugar (e.g., glucosamine, galactosamine etc.) they contain. For example,saccharide moieties may consist of glucuronic acid andacetylgalactosamine (or GlcUA-GalNAc); iduronic acid andacetylgalactosamine (or IdoUA-GlcNAc); glucuronic acid andacetylglucosamine (or GlcUA-GlcNS); or iduronic acid andacetylglucosamine (or IdoUA-GlcNS). In some other cases, saccharidemoieties may consist of galactose and acetylgalactosamine (orGal-GlaNAc).

The saccharide moieties may be processed and functionalized (e.g.,substituted). The saccharide moieties may be substituted by one or moresubstituents which independently are: sulfate, phosphate, carboxylate,acetyl, alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy,halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl,—OR_(a), —SR_(a), —OC(O)R_(a), —N(R_(a))2, —C(O)R_(a), —C(O)OR_(a),—OC(O)N(R_(a))₂, —C(O)N(R_(a))₂, —N(R_(a))C(O)OR_(a),—N(R_(a))C(O)R_(a), —N(R_(a))C(O)N(R_(a))₂, —N(R_(a))C(NR_(a))N(R_(a))₂,—N(R_(a))S(O)_(t)R_(a) (where t is 1 or 2), —S(O)_(t)OR_(a) (where t is1 or 2), —S(O)_(t)N(R_(a))₂ (where t is 1 or 2), or —PO₃(R_(a))₂, whereeach R_(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl. Thefunctionalization of saccharide moieties may be carried our prior to,during, or after the preparation of glycopolymers.

In some cases, the saccharide moieties may be sulfated. The sulfationpattern may be varied. By altering the sulfation pattern of thesaccharide moieties, the molecular recognition and activity ofglycopolymers may be defined and encoded. The precise positioning ofsulfate groups may determine the affinity of glycopolymers for targetmolecules. For example, a single alteration in sulfation pattern (e.g.,removal of a sulfate group from a specific position) may greatly changethe properties of glycopolymers (e.g., abrogate a certain type ofactivity). In some cases, one or more sites of saccharide moieties maybe sulfated, for example, in the case of disaccharide, N-sulfated,2-O-sulfated, 3-O-sulfated, 4-O-sulfated, or 6-O-sulfated. In somecases, to tune the properties of glycopolymers, it may be desirable tohave saccharide moieties with one or more desulfated sites, e.g.,N-desulfated, 2-O-desulfated, 3-O-desulfated, 4-O-desulfated, or6-O-desulfated if a disaccharide moiety is included.

The degree of sulfation may be altered. For example, in some cases, eachsaccharide moiety may consist of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, or moresulfate groups. In one example, each saccharide moiety may consist ofthree sulfate groups. In another example, each saccharide moiety mayconsist of four sulfate groups. In some cases, each saccharide moietymay comprise at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, or moresulfate groups. For example, each saccharide moiety may comprise atleast three or four sulfate groups. In some cases, each saccharidemoiety may comprise no more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60,or more sulfate groups. For example, each saccharide moiety may compriseno more than 10 or 6 sulfate groups. In some cases, the number ofsulfate groups contained in each saccharide moiety falls into a range ofany of the two values described herein.

In some cases, the saccharide moiety may be of the formula:

wherein R₁, R₂, R₃, R₄, R₅, R₆ are each independently hydrogen,hydroxyl, sulfite, sulfate, phosphate, acetyl, or carboxylate.

In some cases, R₁, R₂, R₄, R₅ may each be independently hydrogen,hydroxyl, sulfite, sulfate, phosphate, acetyl, or carboxylate; and R₃and R₆ may each be independently hydroxyl or sulfate. In some cases, R₁,R₂, R₄, R₆ may each be independently hydrogen, hydroxyl, sulfite,sulfate, phosphate, acetyl, or carboxylate; and R₃ and R₅ may each beindependently hydroxyl or sulfate. In some cases, R₁, R₂, R₄ may each beindependently hydrogen, hydroxyl, sulfite, sulfate, phosphate, acetyl,or carboxylate; and R₃, R₅ and R₆ may each be independently hydroxyl orsulfate. In some cases, R₁ and R₂ may each be independently hydrogen,hydroxyl, sulfite, sulfate, phosphate, acetyl, or carboxylate; and R₃,R₄, R₅ and R₆ may each be independently hydroxyl or sulfate. In somecases, R₂ may each be independently hydroxyl, sulfite, sulfate,phosphate, acetyl or carboxylate; and R₁, R₃, R₄, R₅ and R₆ may each beindependently hydroxyl or sulfate. For example, in some cases, R₁, R₃and R₄ may each be independently sulfate; R₂ can be sulfite; and R₅ andR₆ may each be independently hydroxyl. In another example, R₁ and R₄ mayeach be independently sulfate; R₂ can be sulfite; and R₃, R₅ and R₆ mayeach be independently hydroxyl. In a further example, R₁ may each besulfate; R₂ may be sulfite; and R₃, R₄, R₅ and R₆ may each beindependently hydroxyl.

As will be appreciated, in certain cases, it may be desirable to preparea glycopolymer with an indicated percentage of repeating units thatcomprise identical saccharide moieties. For example, in some cases, atleast 80% of the saccharide in the repeating units may comprise aheparan sulfate unit. In some cases, about 1%, 5%, 10%, 15%, 20%, 25%,30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99%, 99.5%, 99.9%, or 100% of the repeating units may compriseidentical saccharide moieties. For example, about 75% or 95% of therepeating units may comprise identical saccharide moieties. In somecases, at least about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%,99.9%, or 100% of the repeating units may comprise identical saccharidemoieties. For example, at least about 50% or 90% of the repeating unitsmay comprise identical saccharide moieties. In some cases, no more thanabout 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or 100% ofthe repeating units may comprise identical saccharide moieties. Forexample, no more than 99% or 90% of the repeating units may compriseidentical saccharide moieties. In some cases, the repeating unitscomprising identical saccharide moieties may constitute a percentage ofthe whole population falling between any of the two values describedherein. For example, about 98.5% of the repeating units may compriseidentical saccharide moieties.

Alternatively, in some cases, a glycopolymer with an indicatedpercentage of repeating units that comprise different saccharidemoieties may be prepared. For example, part of the repeating units maycomprise a disaccharide moiety, while another part of the repeatingunits may comprise a monosaccharide moiety, a trisaccharide moiety, or apentasaccharide moiety. In some cases, about 1%, 5%, 10%, 15%, 20%, 25%,30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99%, 99.5%, 99.9%, or 100% of the repeating units may comprisedifferent saccharide moieties. For example, about 25% or 50% of therepeating units may comprise different saccharide moieties. In somecases, at least about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%,99.9%, or 100% of the repeating units may comprise different saccharidemoieties. For example, at least about 10% or 25% of the repeating unitsmay comprise different saccharide moieties. In some cases, no more thanabout 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or 100% ofthe repeating units may comprise different saccharide moieties. Forexample, no more than about 10% or 25% of the repeating units maycomprise different saccharide moieties. In some cases, the repeatingunits comprising different saccharide moieties may make up a percentageof the whole population falling into a range of any of the two valuesdescribed herein. For example, about 2.5% of the repeating units maycomprise different saccharide moieties.

V. Repeating Units

As described above, each glycopolymer disclosed herein may comprise aplurality of repeating units. Each repeating unit may comprise a polymerbackbone moiety (PB), and/or a linking group (L), and/or a saccharidemoiety (SA). The glycopolymer may comprise a plurality of identical ordifferent repeating units. By “identical repeating units” we mean thateach of the repeating units comprise identical PB, L and SA, which areeach connected in the same configuration. In some cases, theglycopolymers can comprise identical repeating units. In other cases,the glycopolymers can comprise a certain percentage of differingrepeating units (i.e., at least one of PB, L, or SA, is not identicalamong different repeating units).

In some cases, a certain population of the repeating units included in aglycopolymer may be identical. For example, in some cases, equal to orless than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%,55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%of the repeating units in a glycopolymer are identical. In one example,equal to or less than about 75% of the repeating units are identical. Inanother example, equal to or less than about 99% of the repeating unitsare identical. In some cases, more than about 1%, 5%, 10%, 15%, 20%,25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%,95%, 96%, 97%, 98%, 99%, or 100% of the repeating units in aglycopolymer are identical. For example, no more than 90% or 75% of therepeating units are identical. In some cases, the identical repeatingunits included may constitute a certain percentage of the wholepopulation which may fall into a range of any of the two valuesdescribed herein. For example, about 99.5% of the repeating units areidentical.

In some cases, it may be preferred to have glycopolymers with a certainpercentage of differing repeating units such that the properties ofglycopolymers may be tuned. For example, in the case where highlycharged saccharide moieties are attached, to have a less rigid polymerchain, the number or the percentage of the repeating units that comprisesaccharide moieties may be controlled or adjusted (e.g., 80% of therepeating units may comprise saccharide moieties). In some cases, equalto or less than about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or100% of the repeating units in a glycopolymer are different. Forexample, equal to or less than about 25% or 50% of the repeating unitsare different. In some cases, more than about 1%, 5%, 10%, 15%, 20%,25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%,95%, 96%, 97%, 98%, 99%, or 100% of the repeating units in aglycopolymer are different. For example, more than about 10% or 25% ofthe repeating units are different. In some cases, the percentage ofdifferent repeating units may fall between any of the two valuesdescribed herein. For example, about 7.5% of the repeating units aredifferent.

In some cases, the repeating unit may be of the formula:

wherein PB is a polymer backbone moiety and SA is a saccharide moiety;and m is an integer between 1 and 1000.

In some cases, the repeating units may be of the formula:

wherein L is a linking group and SA is a saccharide moiety.

In some cases, the repeating units may be of the formula:

wherein R₁, R₂, R₃ and R₄ may each be independently selected from thegroup consisting of hydrogen, hydroxyl, sulfite, sulfate, acetyl,phosphate and carboxylate; and m is an integer between 1 and 1000. Insome examples, R₃ can be hydroxyl. In other examples, R₃ can be sulfate.

VI. Embodiments/Properties

By utilizing different preparing (e.g., polymerization) conditions, theproperties of glycopolymers can be controlled and tuned. Exemplaryproperties may include, but not limited to the degree of polymerization,length, size, structure, conformation, rigidity, sulfation degree,sulfation pattern, degree of reduction, polydispersity, or combinationsthereof.

The degree of polymerization (DP) may be determined by the number ofrepeating unit, n. In some cases, glycopolymer with a higher DP (i.e.,larger n) may be prepared. In some cases, glycopolymer with a lower DP(i.e., smaller n) may be prepared In some cases, n may be about 1, 2, 5,10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95,100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750,800, 850, 900, 950, 1000, 2000, 3000, 4000, 5000, 6000, 7000, 8000,9000, 10000, 50000, 100000, 500000, or 1000000. For example, n may be15, 30 or 45. In some cases, n may be more than about 1, 2, 5, 10, 15,20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100,150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800,850, 900, 950, 1000, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000,10000, 50000, 100000, 500000, or 1000000. For example, n may be morethan about 10, 30, 100 or 500. In some cases, n may be less than about2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85,90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700,750, 800, 850, 900, 950, 1000, 2000, 3000, 4000, 5000, 6000, 7000, 8000,9000, 10000, 50000, 100000, 500000, or 1000000. For example, n may beless than about 50, 100, 300 or 1000. In some cases, n may fall betweenany two of the values described herein. For example, n may be 46, 48, 99or 155.

The polydispersity index (PDI) or heterogeneity index, is a measure ofthe distribution of molar mass in a given polymer sample, or theuniformity of the given polymer sample. The PDI can be determined by theratio of M_(w)/M_(n), where M_(w) and M_(n) are weight-averaged andnumber-averaged molecular weight (or molar mass), respectively. As thepolymer chains approach uniform chain length, the PDI approaches unity(1). Experimental methods for determining the PDI include massspectrometry, gel permeation chromatography and light scattering.

In many cases, the preparing conditions are used to ensure that thegenerated glycopolymers have a lower PDI, or a narrower distribution.Glycopolymers with a narrow polydispersity (or lower PDI) may be free ofinterfering glycopolymers with differing DP and thus differingbiological effects. For example, in some cases, the PDI may be less thanabout 2. In some cases, a higher PDI may be needed. In some cases, thePDI may be about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0,2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.2, 3.4, 3.6, 3.8,4.0, 4.2, 4.4, 4.6, 4.8, 5.0, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10.For example, the PDI may be about 1.3, 1.4 or 2.0. In some cases, thePDI may be at least about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8,1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.2, 3.4,3.6, 3.8, 4.0, 4.2, 4.4, 4.6, 4.8, 5.0, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9,9.5, 10. For example, the PDI may be at least about 1 or 1.2. In somecases, the PDI may be no more than about 1, 1.1, 1.2, 1.3, 1.4, 1.5,1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9,3.0, 3.2, 3.4, 3.6, 3.8, 4.0, 4.2, 4.4, 4.6, 4.8, 5.0, 5.5, 6, 6.5, 7,7.5, 8, 8.5, 9, 9.5, 10. For example, the PDI may be no more than 2, 3or 4. In some cases, the PDI may fall between any two of the valuesdescribed herein. For example, the PDI may be about 1.25, 1.29 or 1.32.

In some embodiments of the present disclosure, the methods describedherein may provide glycopolymers with substantial homogeneity. Exemplarymethods for purifying the prepared glycopolymers may include but are notlimited to thin layer chromatography; silica gel, reverse phase,ion-exchange, and gel permeation chromatography; and lyopholization. Asused herein, substantial homogeneity means at least about 25% pureincluding about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%,85%, 90%, 95%, 97.5%, 99%, or about 99.5% pure or higher. In some cases,substantially homogeneous may additionally mean having a PDI of lessthan about 5, less than about 4, less than about 3, less than about 2 orless than about 1.5. Methods for determine the purity may include butare not limited to chromatographic methods with the use of refractiveindex and electrochemical detection, light scattering, massspectrometry, and sedimentation velocity. In some cases, a chromophoreor fluorophore may be covalently linked to the glycopolymers of thepresent disclosure allowing detection by spectrophotometric (UVNis) orfluorometric techniques

In some embodiments of the present disclosure, the methods describedherein may provide glycopolymers in high yield. Synthetic yields may becalculated by a number of methods. An exemplary method may comprisesteps: (1) determining the amount and purity of the starting materialand the final product; (2) dividing the pure amount of the final productby that of the starting material; and (3) multiplying the quotientdetermined from step (2) by 100 to obtain a percent yield. To determinethe yield, one may further take into consideration the stoichiometry ofthe reaction. Methods for determining the amount of starting materialand/or final product may include but are not limited to weighing. Insome cases, the final yield may be determined by multiplying the yieldat each step of the synthetic scheme. In some cases, the yield may beabout 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. Forexample, the yield may be about 90%. In some cases, the yield may be atleast about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.For example, the yield may be at least about 75% or 90%. In some cases,the yield may be between any two of the values described herein. Forexample, the yield may be about 91%, 93% or 97%.

Methods for Preparing Glycopolymers

The glycopolymer of the present disclosure can be prepared via a varietyof routes and techniques, such as free radical polymerization,step-growth polymerization, condensation polymerization, cationicaddition polymerization, anionic addition polymerization, livingcationic polymerization, living anionic polymerization, livingring-opening metathesis polymerization, living free radicalpolymerization, reversible addition-fragmentation chain transferpolymerization (RAFT), atom transfer radical polymerization (ATRP),ring-opening polymerization, ring-opening metathesis polymerization(ROMP), emulsion polymerization, solution polymerization, suspensionpolymerization, and precipitation polymerization.

As described elsewhere herein, to temper the polymer properties,different preparing conditions (e.g., temperature, pressure, type andcomposition of initiator and optionally catalyst, type of solvent etc.)may be used.

In some cases, ROMP may be used to prepare the glycopolymers. ROMP is avariant of olefin metathesis and uses optionally substituted cyclicalkyls with at least one carbon-carbon double bond to produce polymersand co-polymers with a low polydispersity index. The mechanism for ROMPfollows similar pathways as olefin metathesis. The catalysts used in theROMP reaction include a wide variety of metals and range from a simpleRuCl₃/alcohol mixture to Grubbs' catalysts (e.g. Grubbs' firstgeneration catalyst, Grubbs' second generation catalyst, etc.).Organometallic catalysts, such as W, Mo, Re, Ru, and Ti carbenescomplexes, may be used. The catalysts used in the ROMP reaction may alsoinclude Schrock catalyst, Grubbs catalyst or derivatives thereof.

ROMP may be applied to strained cyclic monomers. A number of cyclicstructures may be used as monomers in ROMP reaction, such ashomo-monocyclic-, hetero-monocyclic-, homo-bicyclic-, hetero-bicyclic-,homo-tricyclic-, hetero-tricyclic-, homo-polycyclic-,hetero-polycyclic-rings, or derivatives thereof. Non-limiting examplesof monomers may include cyclobutene, cyclopentene, cyclooctene,cyclooctatetraene, norbornene, dicyclopentadiene, or derivativesthereof. In some cases, various substituents can be included in themonomers, in the transition metal catalyst, or in the reaction mixtureto tune the polymerization reaction. Numerous organic and inorganicmoieties may be used as substituents. Non-limiting examples ofsubstituents may include alkyl, heteroalkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl,trifluoromethoxy, nitro, trimethylsilanyl, —OR_(a), —SR_(a),—OC(O)R_(a), —N(R_(a))2, —C(O)R_(a), —C(O)OR_(a), —OC(O)N(R_(a))₂,—C(O)N(R_(a))₂, —N(R_(a))C(O)OR_(a), —N(R_(a))C(O)R_(a),—N(R_(a))C(O)N(R_(a))₂, —N(R_(a))C(NR_(a))N(R_(a))₂,—N(R_(a))S(O)_(t)R_(a) (where t is 1 or 2), —S(O)_(t)OR_(a) (where t is1 or 2), —S(O)_(t)N(R_(a))₂ (where t is 1 or 2), or —PO₃(R_(a))₂, whereeach R_(a) is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl,carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl,heterocycloalkylalkyl, heteroaryl or heteroarylalkyl.

As described elsewhere herein, to prepare glycopolymers with certainproperties, when conducting ROMP reaction, different solvent and/orsolvent systems may be used. The solvent may be polar or non-polar.Non-limiting examples of solvents may include water, methanol, ethanol,isopropanol, dichloromethane, toluene, hexane, tetrachloroethylene,pentane, cyclopentane, cyclohexane, benzene, dioxane, chloroform,diethyl ether, tetrahydrofuran, ethyl acetate, acetone,dimethylformamide, acetonitrile, dimethyl sulfoxide, formic acid,n-Butanol, acetic acid, n-propanol, or combinations thereof. In somecases, a solvent system (or a mix of solvents) may be used. The solventsystem may include two or more different types of solvents, for example,the solvent system may comprise 2, 3, 4, 5 or more different type ofsolvents. In case where a solvent system is used, various ratios ofcomposing solvents may be used. For example, in some cases, a solventsystem may consist of two solvents A and B, where A and B can be any ofthe solvents described above, the ratio of A/B can be about 1:10, 1:9,1:7, 1:6, 1:5, 1:4, 1:3, 1:2, or 1:1. Or, in some cases, the ratio ofA/B may fall into a range of any two of the values described herein.

The preparation of the glycopolymers may comprise one or more steps.Further, one or more types of solvents and/or solvent systems may beused for each single step. The types of solvents and/or solvent systemsused in the preparation process may also vary. In some cases, a singletype of solvent or solvent systems may be used. In some cases, each stepmay involve a distinct type of solvent or solvent system. In some cases,some of the steps may use the same solvent or solvent system, while someother steps may involve the use of differing solvents and/or solventsystems.

It should be noted that, even in cases where the same solvent system(i.e., consisting of same solvent components) is applied, the ratios ofdifferent solvent components may vary, depending upon, the targetproperties of the product (e.g., DP or PDI of the synthesizedglycopolymer). Or, in the cases where more than one step is contained inthe reaction, some certain ratio of solvents in a solvent system may beused in some steps, and another ratio of solvents may be used in othersteps. In some cases, the solvent system may consist of MeOH and(CH₂Cl)₂, and the ratio of MeOH:(CH₂Cl)₂ may be varied from 1:4 to 1:2.5depending upon, the target polymer length. In one example, a ratio of1:4 MeOH:(CH₂Cl)₂ may be used to synthesize polymers with n of 4, 6 and8. In another example, a ratio of 1:3 MeOH:(CH₂Cl)₂ may be used tosynthesize polymers with n of 10 and 15. In a further example, a ratioof 1:2.5 MeOH:(CH₂Cl)₂ may be used for the synthesis of polymers with nof 30 and 50.

As described elsewhere herein, by tuning the reaction conditions, oraltering a number of variables of the reaction, we may adjust andcontrol the properties (e.g., chain length, conformation, configuration,polydispersity, sulfation degree of the saccharide moiety, sulfationpattern of the saccharide moiety etc.) of synthesized glycopolymers.Examples of variables may include, but not limited to, temperature ofthe reaction, amount and type of solvent and/or solvent system, amountand type of catalyst, ratio of solvent system, amount and type ofmonomer, reaction method (e.g., RAFT, ATRP, ROMP etc.), and optionallyamount and type of initiator. As will be appreciated, each of thevariables descried herein may vary among steps, if more than one step isinvolved in the reaction.

Also described in the present disclosure is the modification orfunctionalization of the compound. The compound may comprise the polymerbackbone moiety, the linking group, the saccharide moiety, or theglycopolymer. The modification or functionalization of the compound mayoccur anytime in the process, such as, for example, prior to, during, orafter the preparation of the glycopolymer.

Methods for Assessing Glycopolymer Activity

The activity of the compound provided herein can be assessed by a numberof methods and assays. For example, the biological activity can beassessed in assays which test the activities of saccharide, such aschondroitin sulfate, keratan sulfate, heparin, dermatan sulfate, orheparan sulfate.

In some cases, the anti-inflammatory activity of the compound may beassessed. Methods and assays for testing the anti-inflammatory activitymay include, measuring LPS-induced release of pro-inflammatory cytokinesfrom macrophages (e.g. TNFα, IL-6, IF-γ), or neutrophils (see Examplesbelow). Other relevant assays may comprise effects of lipoteichoic acid,zymosan, DNA, RNA, flagellin or peptidoglycan in the above systems aswell as determination of regulation at the transcriptional level (e.g.Gene-array, qPCR etc). Furthermore, dendritic cell activation oractivation of thrombocytes may also be used as a measure ofanti-inflammatory activity. In some cases, the anti-inflammatoryactivity of the compound may be determined by assays which maycharacterize the binding specificities for chemokines Non-limitingexamples of chemokines may include, but not limited to, CCL2, CCL3,CCL4, CCL5 (or RNATES), CCL7, CCL8, CCL11 (or eotaxin), CCL13, CCL14,CCL19, CCL20, CCL21, CCL22, CCL24, CCL25, CCL26, CCL27, CXCL-8, CXCL10,CXCL12, and CXCL13. In some cases, the tested compound may inhibitRANTES binding by up to about 10%, 20%, 30%, 40%, 50%, 55%, 60%, 65%,70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or100%. In some cases, the compound may inhibit RANTES binding by apercentage between any of the two values described herein, for example,it may be in a range of 10%-100%, 20%-100%, 30%-100%, 40%-100%,50%-100%, 60%-100%, 70%-100%, 80%-100%, or 90%-100%. In some examples,the compound may inhibit RANTES binding by up to about 80% asascertained by an immunosorbent assay. In further examples, the compoundmay inhibit RANTES binding up to about 90% as ascertained by animmunosorbent assay.

In some cases, the compounds may be tested to determine an effective oran inhibitory concentration. In some cases, the compounds may be testedto determine a concentration at which a biological effect is modulatedby a specified percentage relative to a control or a normal protein cellor tissue. For example, the compounds provided herein may be contactedwith neuronal cells at varying concentrations to determine at whatconcentration outgrowth of neurites or axons is affected (e.g. enhancedor inhibited) by a specified percentage such as for example 40%, 50%,60%, 70%, 80%, 90%, 95%, 97.5%, 99%, or 99.5%. Such a concentration istermed the effective concentration or the EC. The EC may further includethe specified percentage. For example, an EC50 is the concentration atwhich a biological effect is modulated (e.g. enhanced or inhibited) by50% by a compound. Similarly an IC50 is the concentration at which abiological effect is inhibited by a compound.

In some cases, the anti-coagulant activity of the compound describedherein may be tested. Such activity may be determined by various methodsand assays, for example, by using the activated partial thromboplastintime (APTT) test, prothrombin time (PT) test or the thrombin clottingtime (TCT) test. An ability to increase the prothrombin time (PT), thethrombin clotting time (TCT) and/or the activated partial thromboplastintime (APTT) may signal the anti-coagulant activity of the testedcompound. In some cases, the APTT of the compound can be equal to ormore than about 0.01 s, 0.05 s, 0.1 s, 0.5 s, 1 s, 5 s, 10 s, 15 s, 20s, 25 s, 30 s, 35 s, 40 s, 45 s, 50 s, 55 s, 60 s, 65 s, 70 s, 75 s, 80s, 85 s, 90 s, 95 s, 100 s, 110 s, 120 s, 130 s, 140 s, 150 s, 160 s,170 s, 180 s, 190 s, 200 s, 225 s, 250 s, 275 s, 300 s, 350 s, 400 s,450 s, 500 s, 550 s, 600 s, 650 s, 700 s, 750 s, 800 s, 850 s, 900 s,950 s or 1000 s, when ascertained by an assay. In some cases, the APTTof the compound may be in a range of any of the two values describedherein. For example, the APTT may be between 0.01 s-1000 s, 0.1 s-1000s, 0.5 s-1000 s, 1 s-1000 s, 5 s-1000 s, 10 s-1000 s, 20 s-1000 s, 50s-1000 s, 100 s-1000 s, 200 s-10005, 300 s-1000 s, 400 s-1000 s, 500s-1000 s, 600 s-1000 s, 700 s-1000 s, 800 s-1000 s, 900 s-1000 s, 0.01s-500 s, 0.1 s-500 s, 0.5 s-500 s, 1 s-500 s, 5 s-500 s, 10 s-500 s, 50s-500 s, 100 s-500 s, 200 s-500 s, 300 s-5005, 400 s-500 s, 0.01 s-250s, 0.1 s-250 s, 0.5 s-250 s, 1 s-250 s, 5 s-250 s, 10 s-250 s, 50 s-250s, 100 s-250 s, 200 s-250 s, 0.01 s-100 s, 0.1 s-100 s, 0.5 s-100 s, 1s-100 s, 5 s-100 s, 10 s-100 s, 25 s-100 s, 50 s-100 s, 75 s-1005, 0.01s-50 s, 0.1 s-50 s, 0.5 s-50 s, 1 s-50 s, 5 s-50 s, 10 s-50 s, 20 s-50s, 30 s-50 s, 40 s-50 s, 0.01 s-25 s, 0.1 s-25 s, 0.5 s-25 s, 1 s-25 s,5 s-25 s, 10 s-25 s, 15 s-25 s, 20 s-25 s, 0.01 s-10 s, 0.1 s-10 s, 0.5s-10 s, 1 s-10 s, 2.5 s-10 s, 5 s-10 s, or 7.5 s-10 s. In some cases,the PT of the compound can be equal to or more than about 0.01 s, 0.05s, 0.1 s, 0.5 s, 1 s, 5 s, 10 s, 15 s, 20 s, 25 s, 30 s, 35 s, 40 s, 45s, 50 s, 55 s, 60 s, 65 s, 70 s, 75 s, 80 s, 85 s, 90 s, 95 s, 100 s,110 s, 120 s, 130 s, 140 s, 150 s, 160 s, 170 s, 180 s, 190 s, 200 s,225 s, 250 s, 275 s, 300 s, 350 s, 400 s, 450 s, 500 s, 550 s, 600 s,650 s, 700 s, 750 s, 800 s, 850 s, 900 s, 950 s or 1000 s, whenascertained by an assay. In some cases, the PT of an effectiveanti-coagulant compound may be in a range of any of the two valuesdescribed herein. For example, the PT may be between 0.01 s-10005, 0.1s-1000 s, 0.5 s-1000 s, 1 s-1000 s, 5 s-1000 s, 10 s-1000 s, 20 s-1000s, 50 s-1000 s, 100 s-1000 s, 200 s-1000 s, 300 s-1000 s, 400 s-1000 s,500 s-1000 s, 600 s-1000 s, 700 s-1000 s, 800 s-1000 s, 900 s-10005,0.01 s-500 s, 0.1 s-500 s, 0.5 s-500 s, 1 s-500 s, 5 s-500 s, 10 s-500s, 50 s-500 s, 100 s-500 s, 200 s-5005, 300 s-500 s, 400 s-500 s, 0.01s-250 s, 0.1 s-250 s, 0.5 s-250 s, 1 s-250 s, 5 s-250 s, 10 s-250 s, 50s-250 s, 100 s-250 s, 200 s-250 s, 0.01 s-100 s, 0.1 s-100 s, 0.5 s-100s, 1 s-100 s, 5 s-100 s, 10 s-100 s, 25 s-100 s, 50 s-1005, 75 s-100 s,0.01 s-50 s, 0.1 s-50 s, 0.5 s-50 s, 1 s-50 s, 5 s-50 s, 10 s-50 s, 20s-50 s, 30 s-50 s, 40 s-50 s, 0.01 s-25 s, 0.1 s-25 s, 0.5 s-25 s, 1s-25 s, 5 s-25 s, 10 s-25 s, 15 s-25 s, 20 s-25 s, 0.01 s-10 s, 0.1 s-10s, 0.5 s-10 s, 1 s-10 s, 2.5 s-10 s, 5 s-10 s, or 7.5 s-10 s. Forexample, when ascertained by an in vitro assay, the prolonged APTTand/or PT with the addition of a compound may indicate itsanti-coagulant activity. In one example, the APTT and PT of the control(i.e., none of the compound is added) may be about 31.2 s and 13.3 s,respectively, when ascertained by an in vitro assay, any compound thatmay elevate the APTT and/or PT (e.g., with APTT longer than 31.2 s andPT longer than 13.3 s) as assessed by the same assay would bear theanticoagulant activity.

In some cases, the anti-coagulant activity of the compound may beassessed by determining its ability to enable the antithrombin III (orATIII) to inhibit the serine proteases Factor Xa (FXa) and FIIa, throughthe binding of the compound to ATIII. FIIa is activated downstream ofFXa in the coagulation cascade, and facilitates blood clotting byconverting soluble fibrinogen to insoluble fibrin strands. By utilizingthis type of assay, the anti-coagulant activity of tested compound maybe quantified by IC50, as described elsewhere herein. The lower the IC50value, the more potent the tested compound may be in reducing the bloodcoagulation. For example, when ascertained by an assay, a control sample(i.e., without the addition of tested compound) may have an anti-FXaIC50 and/or an anti-FIIa IC50 of about 2000 nM or more, and any compoundthat has the ability to lower either one of the IC50s compared to thecontrol may have the anti-coagulant activity. In cases where more thanone tested compound have the anti-coagulant activity, as discussedabove, the one that has the lowest IC50 value may have the highestanti-coagulant activity.

In some cases, the compound may have an anti-FXa IC50 value equal to orless than about 3000 nM, 2000 nM, 1000 nM, 500 nM, 250 nM, 100 nM, 90nM, 80 nM, 70 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 7.5 nM, 5nM, 2.5 nM, 1 nM, 0.5 nM, 0.25 nM, 0.1 nM, 0.05 nM, 0.025 nM, 0.01 nM,0.005 nM, 0.0025 nM, 0.001 nM, 0.0005 nM, 0.00025 nM, 0.0001 nM, 0.00005nM, 0.000025 nM, 0.00001 nM, 0.000005 nM, 0.0000025 nM, or 0.000001 nM,when ascertained by an assay. In some cases, the compound may have ananti-FXa IC50 value between any two of the values described herein, forexample, between 0.000001 nM-3000 nM, 0.000005 nM-3000 nM, 0.00001nM-3000 nM, 0.00005 nM-3000 nM, 0.0001 nM-3000 nM, 0.0005 nM-3000 nM,0.001 nM-3000 nM, 0.005 nM-3000 nM, 0.01 nM-3000 nM, 0.05 nM-3000 nM, 1nM-3000 nM, 10 nM-3000 nM, 50 nM-3000 nM, 100 nM-3000 nM, 250 nM-3000nM, 500 nM-3000 nM, 1000 nM-3000 nM, 2000 nM-3000 nM, 0.000001 nM-2000nM, 0.000005 nM-2000 nM, 0.00001 nM-2000 nM, 0.00005 nM-2000 nM, 0.0001nM-2000 nM, 0.0005 nM-2000 nM, 0.001 nM-2000 nM, 0.005 nM-2000 nM, 0.01nM-2000 nM, 0.05 nM-2000 nM, 1 nM-2000 nM, 10 nM-2000 nM, 50 nM-2000 nM,100 nM-2000 nM, 250 nM-2000 nM, 500 nM-2000 nM, 1000 nM-2000 nM,0.000001 nM-1000 nM, 0.000005 nM-1000 nM, 0.00001 nM-1000 nM, 0.00005nM-1000 nM, 0.0001 nM-1000 nM, 0.0005 nM-1000 nM, 0.001 nM-1000 nM,0.005 nM-1000 nM, 0.01 nM-1000 nM, 0.05 nM-1000 nM, 1 nM-1000 nM, 10nM-1000 nM, 50 nM-1000 nM, 100 nM-1000 nM, 250 nM-1000 nM, 500 nM-1000nM, 750 nM-1000 nM, 0.000001 nM-500 nM, 0.000005 nM-500 nM, 0.00001nM-500 nM, 0.00005 nM-500 nM, 0.0001 nM-500 nM, 0.0005 nM-500 nM, 0.001nM-500 nM, 0.005 nM-500 nM, 0.01 nM-500 nM, 0.05 nM-500 nM, 1 nM-500 nM,10 nM-500 nM, 50 nM-500 nM, 100 nM-500 nM, 250 nM-500 nM, 0.000001nM-250 nM, 0.000005 nM-250 nM, 0.00001 nM-250 nM, 0.00005 nM-250 nM,0.0001 nM-250 nM, 0.0005 nM-250 nM, 0.001 nM-250 nM, 0.005 nM-250 nM,0.01 nM-250 nM, 0.05 nM-250 nM, 1 nM-250 nM, 10 nM-250 nM, 50 nM-250 nM,100 nM-250 nM, 125 nM-250 nM, 0.000001 nM-100 nM, 0.000005 nM-100 nM,0.00001 nM-100 nM, 0.00005 nM-100 nM, 0.0001 nM-100 nM, 0.0005 nM-100nM, 0.001 nM-100 nM, 0.005 nM-100 nM, 0.01 nM-100 nM, 0.05 nM-100 nM, 1nM-100 nM, 10 nM-100 nM, 25 nM-100 nM, 50 nM-100 nM, 75 nM-100 nM, 500nM-0.000001 nM-50 nM, 0.000005 nM-50 nM, 0.00001 nM-50 nM, 0.00005 nM-50nM, 0.0001 nM-50 nM, 0.0005 nM-50 nM, 0.001 nM-50 nM, 0.005 nM-50 nM,0.01 nM-50 nM, 0.05 nM-50 nM, 1 nM-50 nM, 10 nM-50 nM, 25 nM-50 nM,0.000001 nM-10 nM, 0.000005 nM-10 nM, 0.00001 nM-10 nM, 0.00005 nM-10nM, 0.0001 nM-10 nM, 0.0005 nM-10 nM, 0.001 nM-10 nM, 0.005 nM-10 nM,0.01 nM-10 nM, 0.05 nM-10 nM, 1 nM-10 nM, 2.5 nM-10 nM, 5 nM-10 nM, 7.5nM-10 nM, 0.000001 nM-1 nM, 0.000005 nM-1 nM, 0.00001 nM-1 nM, 0.00005nM-1 nM, 0.0001 nM-1 nM, 0.0005 nM-1 nM, 0.001 nM-1 nM, 0.005 nM-1 nM,0.01 nM-1 nM, 0.05 nM-1 nM, 0.1 nM-1 nM, 0.25 nM-1 nM, 0.5 nM-1 nM, or0.75 nM-1 nM.

In some cases, the compound may have an anti-FIIa IC50 value equal to orless than about 3000 nM, 2000 nM, 1000 nM, 500 nM, 250 nM, 100 nM, 90nM, 80 nM, 70 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 7.5 nM, 5nM, 2.5 nM, 1 nM, 0.5 nM, 0.25 nM, 0.1 nM, 0.05 nM, 0.025 nM, 0.01 nM,0.005 nM, 0.0025 nM, 0.001 nM, 0.0005 nM, 0.00025 nM, 0.0001 nM, 0.00005nM, 0.000025 nM, 0.00001 nM, 0.000005 nM, 0.0000025 nM, or 0.000001 nM,when ascertained by an assay. In some cases, the anti-FIIa IC50 of thecompound may be between any of the two values described herein, forexample, between 0.000001 nM-3000 nM, 0.000005 nM-3000 nM, 0.00001nM-3000 nM, 0.00005 nM-3000 nM, 0.0001 nM-3000 nM, 0.0005 nM-3000 nM,0.001 nM-3000 nM, 0.005 nM-3000 nM, 0.01 nM-3000 nM, 0.05 nM-3000 nM, 1nM-3000 nM, 10 nM-3000 nM, 50 nM-3000 nM, 100 nM-3000 nM, 250 nM-3000nM, 500 nM-3000 nM, 1000 nM-3000 nM, 2000 nM-3000 nM, 0.000001 nM-2000nM, 0.000005 nM-2000 nM, 0.00001 nM-2000 nM, 0.00005 nM-2000 nM, 0.0001nM-2000 nM, 0.0005 nM-2000 nM, 0.001 nM-2000 nM, 0.005 nM-2000 nM, 0.01nM-2000 nM, 0.05 nM-2000 nM, 1 nM-2000 nM, 10 nM-2000 nM, 50 nM-2000 nM,100 nM-2000 nM, 250 nM-2000 nM, 500 nM-2000 nM, 1000 nM-2000 nM,0.000001 nM-1000 nM, 0.000005 nM-1000 nM, 0.00001 nM-1000 nM, 0.00005nM-1000 nM, 0.0001 nM-1000 nM, 0.0005 nM-1000 nM, 0.001 nM-1000 nM,0.005 nM-1000 nM, 0.01 nM-1000 nM, 0.05 nM-1000 nM, 1 nM-1000 nM, 10nM-1000 nM, 50 nM-1000 nM, 100 nM-1000 nM, 250 nM-1000 nM, 500 nM-1000nM, 750 nM-1000 nM, 0.000001 nM-500 nM, 0.000005 nM-500 nM, 0.00001nM-500 nM, 0.00005 nM-500 nM, 0.0001 nM-500 nM, 0.0005 nM-500 nM, 0.001nM-500 nM, 0.005 nM-500 nM, 0.01 nM-500 nM, 0.05 nM-500 nM, 1 nM-500 nM,10 nM-500 nM, 50 nM-500 nM, 100 nM-500 nM, 250 nM-500 nM, 0.000001nM-250 nM, 0.000005 nM-250 nM, 0.00001 nM-250 nM, 0.00005 nM-250 nM,0.0001 nM-250 nM, 0.0005 nM-250 nM, 0.001 nM-250 nM, 0.005 nM-250 nM,0.01 nM-250 nM, 0.05 nM-250 nM, 1 nM-250 nM, 10 nM-250 nM, 50 nM-250 nM,100 nM-250 nM, 125 nM-250 nM, 0.000001 nM-100 nM, 0.000005 nM-100 nM,0.00001 nM-100 nM, 0.00005 nM-100 nM, 0.0001 nM-100 nM, 0.0005 nM-100nM, 0.001 nM-100 nM, 0.005 nM-100 nM, 0.01 nM-100 nM, 0.05 nM-100 nM, 1nM-100 nM, 10 nM-100 nM, 25 nM-100 nM, 50 nM-100 nM, 75 nM-100 nM, 500nM-0.000001 nM-50 nM, 0.000005 nM-50 nM, 0.00001 nM-50 nM, 0.00005 nM-50nM, 0.0001 nM-50 nM, 0.0005 nM-50 nM, 0.001 nM-50 nM, 0.005 nM-50 nM,0.01 nM-50 nM, 0.05 nM-50 nM, 1 nM-50 nM, 10 nM-50 nM, 25 nM-50 nM,0.000001 nM-10 nM, 0.000005 nM-10 nM, 0.00001 nM-10 nM, 0.00005 nM-10nM, 0.0001 nM-10 nM, 0.0005 nM-10 nM, 0.001 nM-10 nM, 0.005 nM-10 nM,0.01 nM-10 nM, 0.05 nM-10 nM, 1 nM-10 nM, 2.5 nM-10 nM, 5 nM-10 nM, 7.5nM-10 nM, 0.000001 nM-1 nM, 0.000005 nM-1 nM, 0.00001 nM-1 nM, 0.00005nM-1 nM, 0.0001 nM-1 nM, 0.0005 nM-1 nM, 0.001 nM-1 nM, 0.005 nM-1 nM,0.01 nM-1 nM, 0.05 nM-1 nM, 0.1 nM-1 nM, 0.25 nM-1 nM, 0.5 nM-1 nM, or0.75 nM-1 nM.

Alternative methods may also involve some specific measurements ofprekallikrein activation or the activity of Factor X and othercoagulation factors may be performed to assess the anti-coagulantactivity of the compound. Additionally, peripheral blood mononuclearcells (PBMNC)s can be stimulated by E. coli LPS with or without thecompound and tissue factor and clot formation followed after addition ofhuman plasma, or clotting times for whole blood can be measured. In somecases, the compounds may be tested for their ability to bind proteinsknown to be or suspected of being involved in interactions withnaturally occurring molecules. For example, the compounds may be testedfor their concentration dependent binding abilities and/or specificitiesfor proteins such as chemokines, tumor necrosis factor a, and midkineOther exemplary proteins that bind glycosaminoglycans are provided inthe following table:

TABLE 1 Exemplary Proteins that Bind to GlycosaminoglycansPhysiological/pathophysiological Class Examples effects of bindingmultiple Enzymes glycosaminoglycan biosynthetic enzymes, thrombin andcoagulation factors (proteases), complement proteins (esterases),extracellular superoxide dismutase, topoisomerase Enzyme inhibitorsantithrombin III, heparin cofactor II, coagulation, inflammation,secretory leukocyte proteinase inhibitor, Cl- complement regulationesterase inhibitor Cell adhesion P-selectin, L-selectin, some integrinscell adhesion, inflammation, proteins metastasis Extracellular laminin,fibronectin, collagens, cell adhesion, matrix matrix proteinsthrombospondin, vitionectin, tenascin organization Chemokines plateletfactor IV, ry-interferon, interleukins chemotaxis, signaling,inflammation Growth factors fibroblast growth factors, hepatocyte growthmitogenesis, cell migration factor, vascular endothelial growth factor,insulin-like growth factor-binding proteins, TGF-13-binding proteinsMorphogens hedgehogs, TGF-0 family members cell specification, tissuedifferentiation, development Tyrosine-kinase fibroblast growth factorreceptors, vascular mitogenesis growth factor endothelium growth factorreceptor receptors Lipid-binding apolipoproteins E and B, lipoproteinlipase, lipid metabolism, cell proteins hepatic lipase, annexinsmembrane functions Plaque proteins prion proteins, amyloid proteinplaque formation Nuclear proteins histones, transcription factorsunknown Pathogen surface malaria cirumsporozoite protein pathogeninfections proteins Viral envelope herpes simplex virus, dengue virus,human viral infections proteins immunodeficiency virus, hepatitis CvirusMethods of Treatment

The present disclosure also provides methods for preventing, reducing ortreating a variety of diseases, disorders or conditions in a subject ora biological source (e.g., a blood population) of a subject byadministering the compounds to the subject or the source. In some cases,the methods may comprise administering to the subject a compositioncomprising a substantially homogeneous population of compounds. In somecases, the methods may comprise administering a pharmaceuticalcomposition comprising a therapeutically effective amount of one or moreof the compounds to the subject. The biological source may be in thesubject, or may be isolated from the subject prior to the treatment.

The subject can be diagnosed or be at risk for developing or acquiringan inflammatory condition. The subject can be a human subject.Alternatively, the subject can be a non-human subject, including but notlimited to a non-human primate such as a macaque, chimpanzee, gorilla,vervet, orangutan, baboon or any other non-human primate that can beused as a preclinical model. The non-human subject can be mammal,including but not limited to a mouse, rat, rabbit, pig, sheep, horse,bovine, goat, gerbil, hamster, guinea pig or any other mammal. Thesubject or biological source can be a non-mammalian vertebrate, forexample, a higher vertebrate, or an avian, amphibian or reptilianspecies, or another subject or biological source. In some cases, thesubject can be a transgenic animal. A transgenic animal is a non-humananimal in which one or more of the cells of the animal includes anucleic acid that is non-endogenous (i.e., heterologous) and is presentas an extrachromosomal element in a portion of its cell or stablyintegrated into its germ line DNA (i.e., in the genomic sequence of mostor all of its cells).

The compounds of the present disclosure may be useful for studyingand/or treatment of a number of diseases and conditions. For example, insome cases, the compounds may be useful for the study of the patterningand/or growth of neurons, including neuronal cells from the CNS, brain,spinal cord, and peripheral nerves. In some embodiments, the compoundsmay be useful for the treatment of neurodegenerative diseases, neuraldevelopment disorders, or injuries to neural tissues such as brain orspinal cord injuries. The methods of using the compounds may comprisecontacting a cell or a tissue with a compound provided herein in anamount effective to modulate neuronal outgrowth. In some cases, the cellcan be a neuronal cell. In some cases, the tissue can be a neuronal ornervous tissue. The tissue can be severed or injured. The method may beperformed in vitro or in vivo.

In some cases, the compounds of the present disclosure may be used in anin vitro study of neuronal growth. Any variety of mammalian neuronalcells, including those from the brain, CNS, peripheral nerves and thelike can be treated by the methods provided herein. In addition, thecells may be from any variety of mammalian species such as human, mouse,rat, and any other mammalian species including agricultural stock ornon-domesticated animals, as described elsewhere herein.

In some cases, non-mammalian neuronal cells may be used to screen forcompounds that affect neurite outgrowth, axon guidance, and/or neuralcell viability. For example, teleosts may be used to study the effect ofthe compounds. The use of teleosts in the methods of the presentdisclosure to study neurite outgrowth has several advantages due totheir small size, rapid development, ease of genetic and embryologicalmanipulation, and transparency. The compounds of can be applied toteleosts topically, by injection, or in the liquid medium. The compoundscan be applied to eggs, embryos, or adult teleosts. In some cases, theresults of studies in teleosts may form the basis of future studies inmammalian or human subjects. In other cases, the methods of the presentdisclosure may provide for the study of the compounds in avian subjectssuch as but not limited to the use of a chicken embryo animal model.

In some cases, the compounds may be used to induce neuronal growth incultured neurons, including but not limited to hippocampal neurons,dopaminergic neurons, motor neurons, sensory neurons, and dorsal rootganglion neurons. In certain embodiments, the compounds provided hereinmay be useful for inducing the growth of differentiated neural stemcells prior to implantation. In the case of Parkinson's disease, forinstance, implanted tissue has promise as a replacement for dyingdopaminergic neurons.

In some embodiments, the compounds of the present disclosure may affectaxon guidance, nuerite outgrowth, neural proliferation, and/or neuralcell viability. As neurons begin to assemble into recognizablestructures, they can begin to extend elongated membrane-enclosedprotrusions of cytoplasm that are called processes or neurites. Many ofthese neurites will eventually mature into dendrites or axons. Theseneurites grow toward tissues such as other regions of the nervous systemor other structures on which the neurons will eventually form synapsesor junctions with other tissues such as muscles or glands. These tissuesare often referred to as targets of the neurons, and proper function ofthe nervous system depends on the proper connections between the neuronsand their targets. In some cases, neurite outgrowth is guided byinteraction between the neurons and molecules on the surface of cells orin the extracellular matrix of the tissues through which they grow.These physical interactions attract axons and neurites to grow incertain directions and avoid growing in other directions. In addition,there are diffusible molecules that are similarly attractive orrepellent to neurite and/or axon growth.

Molecules that attract or repel neurite and/or axon growth may includebut not limited to integrins, cadherins, IN-1, laminins, netrins,semaphorins, ephrins, BMPs, Wnts, hedgehog, FGFs, tenascins,proteoglycans, neurotransmitters, nerve growth factor, NCAM, L1, Slitproteins, fibronextin, Comm, Robo, DCC, Robo, Nogo, paxilin, retinoicacid, and glycosaminoglycans. Additionally, there are many knowninhibitors or stimulators of neurite outgrowth that can be used to studyneuronal patterning. Such compounds may include but not limited toBis-I, K252a, okadaic acid, U0126, methyl-murcery, desamethasone,amphetamine, and vincristine. In some cases, anti-proliferativecompounds or neurotoxicants such as, for example, kinase inhibitors,inhibitors of tubulin polymerization, inhibitors of nucleic acidsynthesis, inhibitors of metabolic pathways, cell cycle inhibitors,diphenhydramine, cadmium, lead, 5,5-diphenylhydantoin, and valproic acidcan be used to inhibit neurite outgrowth, neuron proliferation, and/orneuron viability. Other antiproliferative compounds known to affectneural cell proliferation and/or viability may include but not limitedto aphidicolin, hydroxyurea, cytosine arabinoside, 5-flurouracil, andochratoxin A.

The compounds of the present disclosure may be assayed individually fortheir effects on neurite outgrowth, neuron proliferation, and/or neuronviability, or they may be assayed in combination with other compounds oragents described in the present disclosure or in combination with othercompounds known or suspected to affect neurons. The compounds may betested for their ability to augment and/or mitigate the effect of othercompounds on neurons. In some cases, the compounds provided herein maybe administered in combination with tumor necrosis factor a (TNF-a),and/or nerve growth factor (NGF). In some cases, the compounds providedherein may interact with growth factors and cytokines such as but notlimited to TNF-a, FGF, and NGF. In some cases, the compounds providedherein may be tested relative to one or more controls. Controls mayinclude cells not contacted with the compounds of the presentdisclosure, or cells contacted with other compounds. In some cases,controls may be a value understood to be normal, that is not inhibitedor enhanced. For example, neurons under the conditions tested may beknown to exhibit neurite outgrowth to a specified degree or length. Thecompounds may be tested for their ability to induce or inhibit neuriteoutgrowth relative to that specified degree or length (i.e. relative tothe control).

In some embodiments of the present disclosure, the compounds providedherein may be screened for their ability to induce neurite outgrowth.The methods may involve contacting a cultured neuron with a compoundbearing one or more negatively charged groups (e.g. sulfates) anddetermining the increase in neurite length of a treated versus untreatedcontrol. In some cases, a compound of the present disclosure may inhibitneurite outgrowth when applied in solution to the neurons, but mayinduce neurite outgrowth when applied to a substratum. In some cases,the methods and compounds of the present invention may cause an increasein mean neurite length relative to an untreated cell between about 1%and about 50%. In other cases, the increase in mean neurite length isgreater than about 1%, about 2%, about 5%, about 10%, about 15%, about20%, about 25%, about 30%, about 35%, about 40%, about 45%, or greaterthan about 50%. In other cases, the increase in mean neurite length isgreater than 10%, 20%, or 30% relative to an untreated cell.

In some embodiments, the compounds may be screened against neurons thatexhibit various disease phenotypes. For example, superoxide dismutase orSOD1 mutant neurons develop an amyotrophic lateral sclerosis (ALS)-likephenotype. The compounds may be contacted with SOD1 mutant neurons andtested for their ability to inhibit the development of ALS. Similarly,the compounds may be tested against suitable model systems forHuntington's Disease, Parkinson's Disease, and Alzheimer's disease orany other neurodegenerative diseases including but not limited toAlexander's disease, Alper's disease, ataxia telangiectasia, Battendisease, bovine spongiform encephalopathy, Canavan disease, Cockaynesyndrome, corticobasal degeneration, Creutzfeldt-Jakob disease,HIV-associated dementia, Kennedy's disease, Krabbe's disease, lewy bodydementia, Machado-Joseph disease, multiple sclerosis, multiple systematrophy, narcolepsy, neuroborreliosis, Pelizaeus-Merzbacher Disease,peripheral neuropathy, Pick's disease, primary lateral sclerosis, priondiseases, Refsum's disease, Sandhoffs disease, Schilder's disease,subacute combined degeneration of spinal cord secondary to perniciousanaemia, schizophrenia, spinocerebellar ataxia, spinal muscular atrophy,Steele-Richardson-Olszewski disease, and tabes dorsalis.

In some embodiments, the compounds of the present invention may beuseful for the study and/or treatment of other diseases or conditionssuch as but not limited to osteoarthritis, spinal cord injury, neuronalinjury, cancer, blood coagulation diseases or conditions, and deep veinthrombosis. It is known that growth factors involved in cancers interactwith glycosaminoglycans such as but not limited to chondroitin andheparan sulfate. The methods of the present invention provide for theuse of the compounds provided herein for modulation of cancer growthand/or progression.

The methods and compounds of the present disclosure can modulate thegrowth and/or proliferation of cancers, as well as cancer metastasis andangiogenesis, including but not limited to ductal carcinoma in ducttissue in a mammary gland, medullary carcinomas, colloid carcinomas,tubular carcinomas, inflammatory breast cancer; ovarian cancer,including epithelial ovarian tumors such as adenocarcinoma in the ovaryand an adenocarcinoma that has migrated from the ovary into theabdominal cavity, uterine cancer, cervical cancer such as adenocarcinomain the cervix epithelial including squamous cell carcinoma andadenocarcinomas; prostate cancer, such as a prostate cancer selectedfrom the following: an adenocarcinoma or an adenocarinoma that hasmigrated to the bone; pancreatic cancer such as epitheliod carcinoma inthe pancreatic duct tissue and an adenocarcinoma in a pancreatic duct;bladder cancer such as a transitional cell carcinoma in urinary bladder,urothelial carcinomas (transitional cell carcinomas), tumors in theurothelial cells that line the bladder, squamous cell carcinomas,adenocarcinomas, and small cell cancers; leukemia such as acute myeloidleukemia (AML), acute lymphocytic leukemia, chronic lymphocyticleukemia, chronic myeloid leukemia, hairy cell leukemia, myelodysplasia,myeloproliferative disorders, acute myelogenous leukemia (AML), chronicmyelogenous leukemia (CML), mastocytosis, chronic lymphocytic leukemia(CLL), multiple myeloma (MM), and myelodysplastic syndrome (MDS); bonecancer; lung cancer such as non-small cell lung cancer (NSCLC), which isdivided into squamous cell carcinomas, adenocarcinomas, and large cellundifferentiated carcinomas, and small cell lung cancer; skin cancersuch as basal cell carcinoma, melanoma, squamous cell carcinoma andactinic keratosis, which is a skin condition that sometimes developsinto squamous cell carcinoma; eye retinoblastoma; cutaneous orintraocular (eye) melanoma; primary liver cancer (cancer that begins inthe liver); kidney cancer; thyroid cancer such as papillary, follicular,medullary and anaplastic; AIDS-related lymphoma such as diffuse largeB-cell lymphoma, B-cell immunoblastic lymphoma and small non-cleavedcell lymphoma; Kaposi's Sarcoma; viral-induced cancers includinghepatitis B virus (HBV), hepatitis C virus (HCV), and hepatocellularcarcinoma; human lymphotropic virus-type 1 (HTLV-1) and adult T-cellleukemia/lymphoma; and human papilloma virus (HPV) and cervical cancer;central nervous system cancers (CNS) such as primary brain tumor, whichincludes gliomas (astrocytoma, anaplastic astrocytoma, or glioblastomamultiforme), Oligodendroglioma, Ependymoma, Meningioma, Lymphoma,Schwannoma, and Medulloblastoma; peripheral nervous system (PNS) cancerssuch as acoustic neuromas and malignant peripheral nerve sheath tumor(MPNST) including neurofibromas and schwannomas, malignant fibrouscytoma, malignant fibrous histiocytoma, malignant meningioma, malignantmesothelioma, and malignant mixed Mtillerian tumor; oral cavity andoropharyngeal cancer such as, hypopharyngeal cancer, laryngeal cancer,nasopharyngeal cancer, and oropharyngeal cancer; stomach cancer such aslymphomas, gastric stromal tumors, and carcinoid tumors; testicularcancer such as germ cell tumors (GCTs), which include seminomas andnonseminomas, and gonadal stromal tumors, which include Leydig celltumors and Sertoli cell tumors; thymus cancer such as to thymomas,thymic carcinomas, Hodgkin disease, non-Hodgkin lymphomas carcinoids orcarcinoid tumors; rectal cancer; and colon cancer. In some embodiments,the pharmaceutical composition is for the treatment of a non-canceroushyperproliferative disorder such as benign hyperplasia of the skin(e.g., psoriasis), restenosis, or prostate (e.g., benign prostatichypertrophy (BPH)).

In some embodiments of the present disclosure the compounds may inhibitTNF-α and thus reduce inflammation. In some cases, this reduction ininflammation by the administration of compounds of the presentdisclosure may be useful for the treatment of inflammatory diseasesincluding but not limited to chronic inflammatory disease such asarthritis, rheumatoid arthritis, atherosclerosis, and inflammatory boweldisease. Methods for assaying the effect of the compounds of the presentinvention on inflammation may include but not limited to a leukocyteadhesion assay, an NF-κB activation assay, a cytokine release assay, anelastase or tryptase release assay, a reactive oxygen species productionand a reactive nitrogen species production assay.

In some embodiments, the compounds of the present disclosure may beuseful for the enhancement of wound repair. The compounds of the presentdisclosure may be used as mimetics of the naturally occurring moleculesto enhance wound repair. Compounds may be applied topically to a woundor integrated into a bandage or suture material. Methods for assayingwound healing may include but not limited to the scratch assay, theassay described in Rodriguez et al. Methods MOI Biol. 2005; 294:23-9,and the rodent ear-punch assay.

The methods and compounds of the present disclosure can modulate thegrowth and/or proliferation of neoplastic conditions, including but notlimited to Acanthoma, Acinic cell carcinoma, Acoustic neuroma, Acrallentiginous melanoma, Acrospiroma, Acute eosinophilic leukemia, Acutelymphoblastic leukemia, Acute megakaryoblastic leukemia, Acute monocyticleukemia, Acute myeloblastic leukemia with maturation, Acute myeloiddendritic cell leukemia, Acute myeloid leukemia, Acute promyelocyticleukemia, Adamantinoma, Adenocarcinoma, Adenoid cystic carcinoma,Adenoma, Adenomatoid odontogenic tumor, Adrenocortical carcinoma, AdultT-cell leukemia, Aggressive NK-cell leukemia, AIDS-Related Cancers,AIDS-related lymphoma, Alveolar soft part sarcoma, Ameloblastic fibroma,Anal cancer, Anaplastic large cell lymphoma, Anaplastic thyroid cancer,Angioimmunoblastic T-cell lymphoma, Angiomyolipoma, Angiosarcoma,Appendix cancer, Astrocytoma, Atypical teratoid rhabdoid tumor, Basalcell carcinoma, Basal-like carcinoma, B-cell leukemia, B-cell lymphoma,Bellini duct carcinoma, Biliary tract cancer, Bladder cancer, Blastoma,Bone Cancer, Bone tumor, Brain Stem Glioma, Brain Tumor, Breast Cancer,Brenner tumor, Bronchial Tumor, Bronchioloalveolar carcinoma, Browntumor, Burkitt's lymphoma, Cancer of Unknown Primary Site, CarcinoidTumor, Carcinoma, Carcinoma in situ, Carcinoma of the penis, Carcinomaof Unknown Primary Site, Carcinosarcoma, Castleman's Disease, CentralNervous System Embryonal Tumor, Cerebellar Astrocytoma, CerebralAstrocytoma, Cervical Cancer, Cholangiocarcinoma, Chondroma,Chondrosarcoma, Chordoma, Choriocarcinoma, Choroid plexus papilloma,Chronic Lymphocytic Leukemia, Chronic monocytic leukemia, Chronicmyelogenous leukemia, Chronic Myeloproliferative Disorder, Chronicneutrophilic leukemia, Clear-cell tumor, Colon Cancer, Colorectalcancer, Craniopharyngioma, Cutaneous T-cell lymphoma, Degos disease,Dermatofibrosarcoma protuberans, Dermoid cyst, Desmoplastic small roundcell tumor, Diffuse large B cell lymphoma, Dysembryoplasticneuroepithelial tumor, Embryonal carcinoma, Endodermal sinus tumor,Endometrial cancer, Endometrial Uterine Cancer, Endometrioid tumor,Enteropathy-associated T-cell lymphoma, Ependymoblastoma, Ependymoma,Epithelioid sarcoma, Erythroleukemia, Esophageal cancer,Esthesioneuroblastoma, Ewing Family of Tumor, Ewing Family Sarcoma,Ewing's sarcoma, Extracranial Germ Cell Tumor, Extragonadal Germ CellTumor, Extrahepatic Bile Duct Cancer, Extramammary Paget's disease,Fallopian tube cancer, Fetus in fetu, Fibroma, Fibrosarcoma, Follicularlymphoma, Follicular thyroid cancer, Gallbladder Cancer, Gallbladdercancer, Ganglioglioma, Ganglioneuroma, Gastric Cancer, Gastric lymphoma,Gastrointestinal cancer, Gastrointestinal Carcinoid Tumor,Gastrointestinal Stromal Tumor, Gastrointestinal stromal tumor, Germcell tumor, Germinoma, Gestational choriocarcinoma, GestationalTrophoblastic Tumor, Giant cell tumor of bone, Glioblastoma multiforme,Glioma, Gliomatosis cerebri, Glomus tumor, Glucagonoma, Gonadoblastoma,Granulosa cell tumor, Hairy Cell Leukemia, Hairy cell leukemia, Head andNeck Cancer, Head and neck cancer, Heart cancer, Hemangioblastoma,Hemangiopericytoma, Hemangiosarcoma, Hematological malignancy,Hepatocellular carcinoma, Hepatosplenic T-cell lymphoma, Hereditarybreast-ovarian cancer syndrome, Hodgkin Lymphoma, Hodgkin's lymphoma,Hypopharyngeal Cancer, Hypothalamic Glioma, Inflammatory breast cancer,Intraocular Melanoma, Islet cell carcinoma, Islet Cell Tumor, Juvenilemyelomonocytic leukemia, Sarcoma, Kaposi's sarcoma, Kidney Cancer,Klatskin tumor, Krukenberg tumor, Laryngeal Cancer, Laryngeal cancer,Lentigo maligna melanoma, Leukemia, Leukemia, Lip and Oral CavityCancer, Liposarcoma, Lung cancer, Luteoma, Lymphangioma,Lymphangiosarcoma, Lymphoepithelioma, Lymphoid leukemia, Lymphoma,Macroglobulinemia, Malignant Fibrous Histiocytoma, Malignant fibroushistiocytoma, Malignant Fibrous Histiocytoma of Bone, Malignant Glioma,Malignant Mesothelioma, Malignant peripheral nerve sheath tumor,Malignant rhabdoid tumor, Malignant triton tumor, MALT lymphoma, Mantlecell lymphoma, Mast cell leukemia, Mediastinal germ cell tumor,Mediastinal tumor, Medullary thyroid cancer, Medulloblastoma,Medulloblastoma, Medulloepithelioma, Melanoma, Melanoma, Meningioma,Merkel Cell Carcinoma, Mesothelioma, Mesothelioma, Metastatic SquamousNeck Cancer with Occult Primary, Metastatic urothelial carcinoma, MixedMullerian tumor, Monocytic leukemia, Mouth Cancer, Mucinous tumor,Multiple Endocrine Neoplasia Syndrome, Multiple Myeloma, Multiplemyeloma, Mycosis Fungoides, Mycosis fungoides, Myelodysplastic Disease,Myelodysplastic Syndromes, Myeloid leukemia, Myeloid sarcoma,Myeloproliferative Disease, Myxoma, Nasal Cavity Cancer, NasopharyngealCancer, Nasopharyngeal carcinoma, Neoplasm, Neurinoma, Neuroblastoma,Neuroblastoma, Neurofibroma, Neuroma, Nodular melanoma, Non-HodgkinLymphoma, Non-Hodgkin lymphoma, Nonmelanoma Skin Cancer, Non-Small CellLung Cancer, Ocular oncology, Oligoastrocytoma, Oligodendroglioma,Oncocytoma, Optic nerve sheath meningioma, Oral Cancer, Oral cancer,Oropharyngeal Cancer, Osteosarcoma, Osteosarcoma, Ovarian Cancer,Ovarian cancer, Ovarian Epithelial Cancer, Ovarian Germ Cell Tumor,Ovarian Low Malignant Potential Tumor, Paget's disease of the breast,Pancoast tumor, Pancreatic Cancer, Pancreatic cancer, Papillary thyroidcancer, Papillomatosis, Paraganglioma, Paranasal Sinus Cancer,Parathyroid Cancer, Penile Cancer, Perivascular epithelioid cell tumor,Pharyngeal Cancer, Pheochromocytoma, Pineal Parenchymal Tumor ofIntermediate Differentiation, Pineoblastoma, Pituicytoma, Pituitaryadenoma, Pituitary tumor, Plasma Cell Neoplasm, Pleuropulmonaryblastoma, Polyembryoma, Precursor T-lymphoblastic lymphoma, Primarycentral nervous system lymphoma, Primary effusion lymphoma, PrimaryHepatocellular Cancer, Primary Liver Cancer, Primary peritoneal cancer,Primitive neuroectodermal tumor, Prostate cancer, Pseudomyxomaperitonei, Rectal Cancer, Renal cell carcinoma, Respiratory TractCarcinoma Involving the NUT Gene on Chromosome 15, Retinoblastoma,Rhabdomyoma, Rhabdomyosarcoma, Richter's transformation, Sacrococcygealteratoma, Salivary Gland Cancer, Sarcoma, Schwannomatosis, Sebaceousgland carcinoma, Secondary neoplasm, Seminoma, Serous tumor,Sertoli-Leydig cell tumor, Sex cord-stromal tumor, Sezary Syndrome,Signet ring cell carcinoma, Skin Cancer, Small blue round cell tumor,Small cell carcinoma, Small Cell Lung Cancer, Small cell lymphoma, Smallintestine cancer, Soft tissue sarcoma, Somatostatinoma, Soot wart,Spinal Cord Tumor, Spinal tumor, Splenic marginal zone lymphoma,Squamous cell carcinoma, Stomach cancer, Superficial spreading melanoma,Supratentorial Primitive Neuroectodermal Tumor, Surfaceepithelial-stromal tumor, Synovial sarcoma, T-cell acute lymphoblasticleukemia, T-cell large granular lymphocyte leukemia, T-cell leukemia,T-cell lymphoma, T-cell prolymphocytic leukemia, Teratoma, Terminallymphatic cancer, Testicular cancer, Thecoma, Throat Cancer, ThymicCarcinoma, Thymoma, Thyroid cancer, Transitional Cell Cancer of RenalPelvis and Ureter, Transitional cell carcinoma, Urachal cancer, Urethralcancer, Urogenital neoplasm, Uterine sarcoma, Uveal melanoma, VaginalCancer, Verner Morrison syndrome, Verrucous carcinoma, Visual PathwayGlioma, Vulvar Cancer, Waldenstrom's macroglobulinemia, Warthin's tumor,Wilms' tumor, or any combination thereof.

In some cases, the methods may involve the treatment of solid tumors.Solid tumors may include malignancies (e.g., sarcomas, adenocarcinomas,and carcinomas) of the various organ systems, such as those of lung,breast, lymphoid, gastrointestinal (e.g., colon), and genitourinary(e.g., renal, urothelial, or testicular tumors) tracts, pharynx,prostate, and ovary. Exemplary adenocarcinomas may include colorectalcancers, renal-cell carcinoma, liver cancer, non-small cell carcinoma ofthe lung, and cancer of the small intestine. Additional exemplary solidtumors may include: fibrosarcoma, myxosarcoma, liposarcoma,chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma,endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma,synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma,rhabdomyosarcoma, gastrointestinal system carcinomas, colon carcinoma,pancreatic cancer, breast cancer, genitourinary system carcinomas,ovarian cancer, prostate cancer, squamous cell carcinoma, basal cellcarcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous glandcarcinoma, papillary carcinoma, papillary adenocarcinomas,cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renalcell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma,seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, endocrinesystem carcinomas, testicular tumor, lung carcinoma, small cell lungcarcinoma, non-small cell lung carcinoma, bladder carcinoma, epithelialcarcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma,ependymoma, pinealoma, hemangioblastoma, acoustic neuroma,oligodendroglioma, meningioma, melanoma, neuroblastoma, andretinoblastoma.

In some cases, the methods may be used for detecting, preventing,reducing or treating coagulation disorders in the subject. Coagulationdisorders deal with disruption of the body's ability to control bloodclotting. Non-limiting examples of coagulation disorders may includeHemophilia, Hemophilia A, Hemophilia B, Hemophilia C, Christmas disease,Factor IX deficiency, Disseminated intravascular coagulation disorder,consumption coagulopathy, Thrombocytopenia, Von Willebrand's disease,Hypoprothrombinemia, Factor XI deficiency, Factor VII deficiency, andserum prothrombin conversion accelerator (SPCA) deficiency.

In some cases, the methods may comprise the treatment of autoimmunedisorders. Examples of autoimmune disorders may include, but not limitedto, Crohn's disease, ulcerative colitis, psoriasis, psoriatic arthritis,juvenile arthritis and ankylosing spondilitis, Other non-limitingexamples of autoimmune disorders include autoimmune diabetes, multiplesclerosis, systemic lupus erythematosus (SLE), rheumatoid spondylitis,gouty arthritis, allergy, autoimmune uveitis, nephrotic syndrome,multisystem autoimmune diseases, autoimmune hearing loss, adultrespiratory distress syndrome, shock lung, chronic pulmonaryinflammatory disease, pulmonary sarcoidosis, pulmonary fibrosis,silicosis, idiopathic interstitial lung disease, chronic obstructivepulmonary disease, asthma, restenosis, spondyloarthropathies, Reiter'ssyndrome, autoimmune hepatitis, inflammatory skin disorders, vasculitisoflarge vessels, medium vessels or small vessels, endometriosis,prostatitis and Sjogren's syndrome. Undesirable immune response can alsobe associated with or result in, e.g., asthma, emphysema, bronchitis,psoriasis, allergy, anaphylaxsis, auto-immune diseases, rhuematoidarthritis, graft versus host disease, transplantation rejection, lunginjuries, and lupus erythematosus. The pharmaceutical compositions ofthe present invention can be used to treat other respiratory diseasesincluding but not limited to diseases affecting the lobes of lung,pleural cavity, bronchial tubes, trachea, upper respiratory tract, orthe nerves and muscle for breathing. The methods of the invention can befurther used to treat multiorgan failure.

In some cases, the methods can also be used for the treatment ofdisorders involving platelet aggregation or platelet adhesion, includingbut not limited to Bernard-Soulier syndrome, Glanzmann's thrombasthenia,Scott's syndrome, von Willebrand disease, Hermansky-Pudlak Syndrome, andGray platelet syndrome.

In some cases, the compounds, compositions or pharmaceuticalcompositions may be used to prevent, reduce or treat an inflammatorycondition. The inflammatory condition may be an acute systemicinflammatory disease or a chronic inflammatory disease. Examples ofinflammatory conditions may include but not limited to systemicinflammatory response syndrome (SIRS), ARDS, sepsis, inflammatory boweldisease, inflammatory skin diseases, psoriasis, eczema, sclerodermasevere sepsis, septic shock erysipelas, meningitis, arthritis,rheumatoid arthritis, toxic shock syndrome, diverticulitis,appendicitis, pancreatitis, cholecystitis, colitis, cellulitis, burnwound infections, pneumonia, urinary tract infections, postoperativeinfections, peritonitis cystic fibrosis, COPD and other pulmonarydiseases, gastrointestinal disease including chronic skin and stomachulcerations, atopic dermatitis, oral ulcerations, aphtous ulcers,genital ulcerations and inflammatory changes, parodontitis, eyeinflammations including conjunctivitis and keratitis, external otitis,mediaotitis and genitourinary inflammations.

In some cases, the methods can be used for treating liver diseases(including diabetes), pancreatitis or kidney disease (includingproliferative glomerulonephritis and diabetes-induced renal disease) orpain, neurological or neurodegenerative diseases (including, but notlimited to, Alzheimer's disease, Huntington's disease, CNS trauma, andstroke).

In some cases, the methods can be used for the prevention of blastocyteimplantation in a mammal skeletal muscle atrophy, skeletal musclehypertrophy, leukocyte recruitment in cancer tissue, invasionmetastasis, melanoma, sarcoma, acute and chronic bacterial and viralinfections, sepsis, glomerulo sclerosis, glomerulo, nephritis, orprogressive renal fibrosis.

As will be appreciated, compounds described in the present disclosuremay be applied for the prevention and treatment for a specific type ofdisease (or condition) as noted above, or they may be used to prevent ortreat a combination of diseases or conditions described in the presentdisclosure. For example, in some cases, the compounds may be used totreat only the inflammatory conditions in a subject. In some cases, thecompounds may only be applicable in reducing the coagulation in a bloodpopulation of a subject. In some cases, the compounds disclosed hereinmay be useful both in treating the inflammatory conditions in a subjectand reducing the coagulation in a blood population of a subject.

Compositions

I. General

One aspect of the present disclosure provides a composition comprising asubstantially homogeneous population of one or more compounds disclosedelsewhere herein. By “substantially homogeneous” we mean the content orthe concentration of compounds included in the composition exceeds acertain limit, for example, more than 50% (w/w).

In some cases, the concentration of one or more of the compounds may beabout 100%, 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%,87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 75%, 70%, 65%, 60%, 55%, 50%,45%, 40%, 35%, 30%, 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%,11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%,0.1%, 0.09%, 0.08%, 0.07%, 0:06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%,0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%,0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%,or 0.0001% w/w, w/v or v/v. For example, the concentration of one ormore of the compounds may be about 50%, 75%, 90% or 99% w/w, w/v or v/v.

In some cases, the concentration of one or more of the compounds may beless than about 100%, 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%,89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 75%, 70%, 65%, 60%,55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%,13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%,0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0:06%, 0.05%, 0.04%, 0.03%,0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%,0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%,0.0003%, 0.0002%, or 0.0001% w/w, w/v or v/v. For example, theconcentration of one or more of the compounds may be less than about100%, 90%, 75% or 50% w/w, w/v or v/v.

In some case, the concentration of one or more of the compounds may begreater than 99.9999%, 99.999%, 99.99%, 99.9%, 99%, 98%, 97%, 96%, 95%,94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%,80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%,10%, 5%, 2.5%, 1%, 0.5%, 0.25%, 0.1%, 0.05%, 0.025%, 0.01%, 0.005%,0.0025%, 0.001%, 0.0005%, 0.00025%, or 0.0001% w/w, w/v, or v/v. Forexample, the concentration of one or more of the compounds may begreater than about 50%, 75%, 95% or 99% w/w, w/v or v/v.

In some cases, the concentration of one or more of the compounds may bein a range of any of the two values described herein. For example, theconcentration may be from approximately 0.0001% to 100%, approximately0.0001% to 90%, approximately 0.0001% to 80%, approximately 0.0001% to70%, approximately 0.0001% to 60%, approximately 0.0001% to 50%,approximately 0.0001% to 40%, approximately 0.0001% to 30%,approximately 0.0001% to 20%, approximately 0.0001% to 10%,approximately 0.0001% to 5%, approximately 0.0001% to 2.5%,approximately 0.0001% to 1%, approximately 0.001% to 100%, approximately0.001% to 90%, approximately 0.001% to 80%, approximately 0.001% to 70%,approximately 0.001% to 60%, approximately 0.001% to 50%, approximately0.001% to 40%, approximately 0.001% to 30%, approximately 0.001% to 20%,approximately 0.001% to 10%, approximately 0.001% to 5%, approximately0.001% to 2.5%, approximately 0.001% to 1%, approximately 0.01% to 100%,approximately 0.01% to 90%, approximately 0.01% to 80%, approximately0.01% to 70%, approximately 0.01% to 60%, approximately 0.01% to 50%,approximately 0.01% to 40%, approximately 0.01% to 30%, approximately0.01% to 20%, approximately 0.01% to 10%, approximately 0.01% to 5%,approximately 0.01% to 2.5%, approximately 0.01% to 1%, approximately0.1% to 100%, approximately 0.1% to 90%, approximately 0.1% to 80%,approximately 0.1% to 70%, approximately 0.1% to 60%, approximately 0.1%to 50%, approximately 0.1% to 40%, approximately 0.1% to 30%,approximately 0.1% to 20%, approximately 0.1% to 10%, approximately 0.1%to 5%, approximately 0.1% to 2.5%, approximately 0.1% to 1%,approximately 1% to 100%, approximately 1% to 90%, approximately 1% to80%, approximately 1% to 70%, approximately 1% to 60%, approximately 1%to 50%, approximately 1% to 40%, approximately 1% to 30%, approximately1% to 20%, approximately 1% to 10%, approximately 1% to 5%,approximately 1% to 2.5 w/w, w/v or v/v.

The quantity of one or more of the compounds included in the compositionmay vary. In some cases, a large quantity of compounds may be used. Insome cases, a small quantity of compounds may be used. In some cases,the quantity of compounds contained in the composition may be equal toor less than about 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0 g, 7.5 g, 7.0 g, 6.5g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g, 3.0 g, 2.5 g, 2.0 g, 1.5 g,1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g, 0.65 g, 0.6 g, 0.55g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, 0.3 g, 0.25 g, 0.2 g, 0.15 g, 0.1 g,0.09 g, 0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g, 0.03 g, 0.02 g, 0.01 g,0.009 g, 0.008 g, 0.007 g, 0.006 g, 0.005 g, 0.004 g, 0.003 g, 0.002 g,0.001 g, 0.0009 g, 0.0008 g, 0.0007 g, 0.0006 g, 0.0005 g, 0.0004 g,0.0003 g, 0.0002 g, or 0.0001 g. For example, the quantity of one ormore of the compounds included in may be equal to or less than about 5g, 1 g, 0.5 g or 0.05 g.

In some cases, the quantity of compounds in the composition may be morethan about 0.0001 g, 0.0002 g, 0.0003 g, 0.0004 g, 0.0005 g, 0.0006 g,0.0007 g, 0.0008 g, 0.0009 g, 0.001 g, 0.0015 g, 0.002 g, 0.0025 g,0.003 g, 0.0035 g, 0.004 g, 0.0045 g, 0.005 g, 0.0055 g, 0.006 g, 0.0065g, 0.007 g, 0.0075 g, 0.008 g, 0.0085 g, 0.009 g, 0.0095 g, 0.01 g,0.015 g, 0.02 g, 0.025 g, 0.03 g, 0.035 g, 0.04 g, 0.045 g, 0.05 g,0.055 g, 0.06 g, 0.065 g, 0.07 g, 0.075 g, 0.08 g, 0.085 g, 0.09 g,0.095 g, 0.1 g, 0.15 g, 0.2 g, 0.25 g, 0.3 g, 0.35 g, 0.4 g, 0.45 g, 0.5g, 0.55 g, 0.6 g, 0.65 g, 0.7 g, 0.75 g, 0.8 g, 0.85 g, 0.9 g, 0.95 g, 1g, 1.5 g, 2 g, 2.5, 3 g, 3.5, 4 g, 4.5 g, 5 g, 5.5 g, 6 g, 6.5 g, 7 g,7.5 g, 8 g, 8.5 g, 9 g, 9.5 g, or 10 g. For example, the quantity of oneor more of the compounds in the composition may be more than about0.0001 g, 0.05 g, 1 g or 2.5 g.

In some cases, the quantity of compounds in the composition may be in arange of any two of the values described herein. For example, acomposition may comprise one or more of the compounds whose quantity isin the range of 0.0001-10 g, 0.0001-9 g, 0.0001-8 g, 0.0001-7 g,0.0001-6 g, 0.0001-5 g, 0.0001-4 g, 0.0001-3 g, 0.0001-2 g, 0.0001-1 g,0.0005-10 g, 0.0005-9 g, 0.0005-8 g, 0.0005-7 g, 0.0005-6 g, 0.0005-5 g,0.0005-4 g, 0.0005-3 g, 0.0005-2 g, 0.0005-1 g, 0.001-10 g, 0.001-9 g,0.001-8 g, 0.001-7 g, 0.001-6 g, 0.001-5 g, 0.001-4 g, 0.001-3 g,0.001-2 g, 0.001-1 g, 0.005-10 g, 0.005-9 g, 0.005-8 g, 0.005-7 g,0.005-6 g, 0.005-5 g, 0.005-4 g, 0.005-3 g, 0.005-2 g, 0.005-1 g,0.01-10 g, 0.01-9 g, 0.01-8 g, 0.01-7 g, 0.01-6 g, 0.01-5 g, 0.01-4 g,0.01-3 g, 0.01-2 g, 0.01-1 g, 0.05-10 g, 0.05-9 g, 0.05-8 g, 0.05-7 g,0.05-6 g, 0.05-5 g, 0.05-4 g, 0.05-3 g, 0.05-2 g, 0.05-1 g, 0.1-10 g,0.1-9 g, 0.1-8 g, 0.1-7 g, 0.1-6 g, 0.1-5 g, 0.1-4 g, 0.1-3 g, 0.1-2 g,0.1-1 g, 0.5-10 g, 0.5-9 g, 0.1-8 g, 0.1-7 g, 0.1-6 g, 0.1-5 g, 0.1-4 g,0.1-3 g, 0.1-2 g, 0.1-1 g, 0.5-10 g, 0.5-9 g, 0.5-8 g, 0.5-7 g, 0.5-6 g,0.5-5 g, 0.5-4 g, 0.5-3 g, 0.5-2 g, 0.5-1 g, 1-10 g, 1-9 g, 1-8 g, 1-7g, 1-6 g, 1-5 g, 1-4 g, 1-3 g, or 1-2 g.

II. Pharmaceutical Compositions and Methods of Administration

The present disclosure also provides pharmaceutical compositions. Thepharmaceutical compositions are typically formulated to provide one ormore compounds of the present disclosure as the active ingredient, or apharmaceutically acceptable salt, ester, prodrug, solvate, hydrate orderivative thereof. The pharmaceutical composition may comprise atherapeutically effective amount of the compound of the presentdisclosure. Where desired, the pharmaceutical compositions containpharmaceutically acceptable salt and/or coordination complex thereof,and one or more pharmaceutically acceptable excipients, carriers,including inert solid diluents and fillers, diluents, including sterileaqueous solution and various organic solvents, permeation enhancers,solubilizers and adjuvants.

The pharmaceutical compositions can be administered alone or incombination with one or more other agents, which are also typicallyadministered in the form of pharmaceutical compositions. In some cases,it may be desirable that the one or more of the compounds in the presentdisclosure and other agent(s) be mixed into a preparation or allcomponents may be formulated into separate preparations to use them incombination separately or at the same time.

As will be appreciated, as the active ingredient, the concentration ofone or more of the compounds contained in pharmaceutical compositionsmay vary. In some cases, a high concentration of compounds may beincluded. In some cases, a low concentration of compounds may be used.In some cases, the concentration of one or more of the compounds may beequal to or less than about 100%, 99%, 98%, 97%, 96%, 95%, 94%, 93%,92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 75%,70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 19%, 18%, 17%,16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%,0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0:06%, 0.05%, 0.04%,0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%,0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%,0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w, w/v or v/v. For example, theconcentration of one or more of the compounds may be equal to or lessthan about 50%, 75%, 90% or 99% w/w, w/v or v/v.

In some case, the concentration of one or more of the compounds may begreater than 99.9999%, 99.999%, 99.99%, 99.9%, 99%, 98.5%, 98%, 97.5%,97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%,83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 70%, 65%, 60%, 55%, 50%,45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 2.5%, 1%, 0.5%, 0.25%, 0.1%,0.05%, 0.025%, 0.01%, 0.005%, 0.0025%, 0.001%, 0.0005%, 0.00025%, or0.0001% w/w, w/v, or v/v. For example, the concentration of one or moreof the compounds may be greater than about 50%, 75%, 95% or 99% w/w, w/vor v/v.

In some cases, the concentration of one or more of the compounds may bein a range of any of the two values described herein. For example, theconcentration may be from approximately 0.0001% to 100%, approximately0.0001% to 90%, approximately 0.0001% to 80%, approximately 0.0001% to70%, approximately 0.0001% to 60%, approximately 0.0001% to 50%,approximately 0.0001% to 40%, approximately 0.0001% to 30%,approximately 0.0001% to 20%, approximately 0.0001% to 10%,approximately 0.0001% to 5%, approximately 0.0001% to 2.5%,approximately 0.0001% to 1%, approximately 0.001% to 100%, approximately0.001% to 90%, approximately 0.001% to 80%, approximately 0.001% to 70%,approximately 0.001% to 60%, approximately 0.001% to 50%, approximately0.001% to 40%, approximately 0.001% to 30%, approximately 0.001% to 20%,approximately 0.001% to 10%, approximately 0.001% to 5%, approximately0.001% to 2.5%, approximately 0.001% to 1%, approximately 0.01% to 100%,approximately 0.01% to 90%, approximately 0.01% to 80%, approximately0.01% to 70%, approximately 0.01% to 60%, approximately 0.01% to 50%,approximately 0.01% to 40%, approximately 0.01% to 30%, approximately0.01% to 20%, approximately 0.01% to 10%, approximately 0.01% to 5%,approximately 0.01% to 2.5%, approximately 0.01% to 1%, approximately0.1% to 100%, approximately 0.1% to 90%, approximately 0.1% to 80%,approximately 0.1% to 70%, approximately 0.1% to 60%, approximately 0.1%to 50%, approximately 0.1% to 40%, approximately 0.1% to 30%,approximately 0.1% to 20%, approximately 0.1% to 10%, approximately 0.1%to 5%, approximately 0.1% to 2.5%, approximately 0.1% to 1%,approximately 1% to 100%, approximately 1% to 90%, approximately 1% to80%, approximately 1% to 70%, approximately 1% to 60%, approximately 1%to 50%, approximately 1% to 40%, approximately 1% to 30%, approximately1% to 20%, approximately 1% to 10%, approximately 1% to 5%,approximately 1% to 2.5% w/w, w/v or v/v.

The quantity of one or more of the compounds included in pharmaceuticalcompositions may vary. In some cases, a large quantity of compounds maybe used. In some cases, a small quantity of compounds may be used. Insome cases, the quantity of compounds contained in pharmaceuticalcompositions may be equal to or less than about 10 g, 9.5 g, 9.0 g, 8.5g, 8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g,3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g,0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, 0.3 g, 0.25g, 0.2 g, 0.15 g, 0.1 g, 0.09 g, 0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g,0.03 g, 0.02 g, 0.01 g, 0.009 g, 0.008 g, 0.007 g, 0.006 g, 0.005 g,0.004 g, 0.003 g, 0.002 g, 0.001 g, 0.0009 g, 0.0008 g, 0.0007 g, 0.0006g, 0.0005 g, 0.0004 g, 0.0003 g, 0.0002 g, or 0.0001 g. For example, thequantity of one or more of the compounds contained in pharmaceuticalcompositions may be equal to or less than about 5 g, 1 g, 0.5 g or 0.05g.

In some cases, the quantity of compounds contained in pharmaceuticalcompositions may be greater than about 0.0001 g, 0.0002 g, 0.0003 g,0.0004 g, 0.0005 g, 0.0006 g, 0.0007 g, 0.0008 g, 0.0009 g, 0.001 g,0.0015 g, 0.002 g, 0.0025 g, 0.003 g, 0.0035 g, 0.004 g, 0.0045 g, 0.005g, 0.0055 g, 0.006 g, 0.0065 g, 0.007 g, 0.0075 g, 0.008 g, 0.0085 g,0.009 g, 0.0095 g, 0.01 g, 0.015 g, 0.02 g, 0.025 g, 0.03 g, 0.035 g,0.04 g, 0.045 g, 0.05 g, 0.055 g, 0.06 g, 0.065 g, 0.07 g, 0.075 g, 0.08g, 0.085 g, 0.09 g, 0.095 g, 0.1 g, 0.15 g, 0.2 g, 0.25 g, 0.3 g, 0.35g, 0.4 g, 0.45 g, 0.5 g, 0.55 g, 0.6 g, 0.65 g, 0.7 g, 0.75 g, 0.8 g,0.85 g, 0.9 g, 0.95 g, 1 g, 1.5 g, 2 g, 2.5, 3 g, 3.5 g, 4 g, 4.5 g, 5g, 5.5 g, 6 g, 6.5 g, 7 g, 7.5 g, 8 g, 8.5 g, 9 g, 9.5 g, or 10 g. Forexample, the quantity of one or more of the compounds contained inpharmaceutical compositions may be greater than about 0.0001 g, 0.05 g,1 g or 2.5 g.

In some cases, the quantity of compounds in pharmaceutical compositionsmay be in a range of any two of the values described herein. Forexample, a pharmaceutical composition may comprise one or more of thecompounds whose quantity is in the range of 0.0001-10 g, 0.0001-9 g,0.0001-8 g, 0.0001-7 g, 0.0001-6 g, 0.0001-5 g, 0.0001-4 g, 0.0001-3 g,0.0001-2 g, 0.0001-1 g, 0.0005-10 g, 0.0005-9 g, 0.0005-8 g, 0.0005-7 g,0.0005-6 g, 0.0005-5 g, 0.0005-4 g, 0.0005-3 g, 0.0005-2 g, 0.0005-1 g,0.001-10 g, 0.001-9 g, 0.001-8 g, 0.001-7 g, 0.001-6 g, 0.001-5 g,0.001-4 g, 0.001-3 g, 0.001-2 g, 0.001-1 g, 0.005-10 g, 0.005-9 g,0.005-8 g, 0.005-7 g, 0.005-6 g, 0.005-5 g, 0.005-4 g, 0.005-3 g,0.005-2 g, 0.005-1 g, 0.01-10 g, 0.01-9 g, 0.01-8 g, 0.01-7 g, 0.01-6 g,0.01-5 g, 0.01-4 g, 0.01-3 g, 0.01-2 g, 0.01-1 g, 0.05-10 g, 0.05-9 g,0.05-8 g, 0.05-7 g, 0.05-6 g, 0.05-5 g, 0.05-4 g, 0.05-3 g, 0.05-2 g,0.05-1 g, 0.1-10 g, 0.1-9 g, 0.1-8 g, 0.1-7 g, 0.1-6 g, 0.1-5 g, 0.1-4g, 0.1-3 g, 0.1-2 g, 0.1-1 g, 0.5-10 g, 0.5-9 g, 0.1-8 g, 0.1-7 g, 0.1-6g, 0.1-5 g, 0.1-4 g, 0.1-3 g, 0.1-2 g, 0.1-1 g, 0.5-10 g, 0.5-9 g, 0.5-8g, 0.5-7 g, 0.5-6 g, 0.5-5 g, 0.5-4 g, 0.5-3 g, 0.5-2 g, 0.5-1 g, 1-10g, 1-9 g, 1-8 g, 1-7 g, 1-6 g, 1-5 g, 1-4 g, 1-3 g, or 1-2 g.

The pharmaceutical composition according to the disclosure may beeffective over a wide dosage range. For example, in the treatment ofadult humans, dosages from about 0.01 to about 1000 mg, from about 0.5to about 100 mg, from about 1 to about 50 mg per day, and from about 5to about 40 mg per day may be used. The exact dosage may depend upon,for example, the route of administration, the form in which the compoundis administered, the subject to be treated, the body weight of thesubject to be treated, and the preference and experience of theattending physician.

Provided in the present disclosure are a number of methods ofadministering pharmaceutical compositions described elsewhere herein tothe subjects. Described below are non-limiting exemplary pharmaceuticalcompositions and methods for preparing the same.

A. Pharmaceutical Compositions for Oral Administration

In some embodiments, the invention provides a pharmaceutical compositionfor oral administration containing a compound of the disclosure, and apharmaceutical excipient suitable for oral administration.

In some embodiments, the invention provides a solid pharmaceuticalcomposition for oral administration comprising: (i) an effective amountof a compound of the disclosure; (ii) an effective amount of a secondagent; and/or (iii) a pharmaceutical excipient suitable for oraladministration. In some embodiments, the composition may furthercontain: (iv) an effective amount one or more of additional agents,e.g., a third agent.

In some embodiments, the pharmaceutical composition may be a liquidpharmaceutical composition suitable for oral consumption. Pharmaceuticalcompositions of the disclosure for oral administration can be presentedas discrete dosage forms, such as capsules, cachets, or tablets, orliquids or aerosol sprays each containing a predetermined amount of anactive ingredient as a powder or in granules, a solution, or asuspension in an aqueous or non-aqueous liquid, an oil-in-wateremulsion, or a water-in-oil liquid emulsion. Such dosage forms can beprepared by a number of methods of pharmacy, but most, if not allmethods may include the step of bringing the active ingredient intoassociation with the carrier, which may constitute one or more necessaryingredients. In general, the compositions may be prepared by uniformlyand intimately admixing the active ingredient with liquid carriers orfinely divided solid carriers or both, and then, if necessary, shapingthe product into the desired presentation. For example, a tablet can beprepared by compression or molding, optionally with one or moreaccessory ingredients. Compressed tablets can be prepared by compressingin a suitable machine the active ingredient in a free-flowing form suchas powder or granules, optionally mixed with an excipient such as, butnot limited to, a binder, a lubricant, an inert diluent, and/or asurface active or dispersing agent. Molded tablets can be made bymolding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

The present disclosure further encompasses anhydrous pharmaceuticalcompositions and dosage forms comprising one or more active ingredients,since water can facilitate the degradation of some ingredients. Forexample, water may be added (e.g., 5%) in the pharmaceutical arts as amethod of simulating long-term storage in order to determinecharacteristics such as shelf-life or the stability of formulations overtime Anhydrous pharmaceutical compositions and dosage forms of thedisclosure can be prepared using anhydrous or low moisture containingingredients and low moisture or low humidity conditions. Pharmaceuticalcompositions and dosage forms of the disclosure which may containlactose can be made anhydrous if substantial contact with moistureand/or humidity during manufacturing, packaging, and/or storage isexpected. An anhydrous pharmaceutical composition may be prepared andstored such that its anhydrous nature is maintained. Accordingly,anhydrous compositions may be packaged using a number of materials toprevent exposure to water such that they can be included in suitableformulary kits. Examples of suitable packaging may include, but are notlimited to, hermetically sealed foils, plastic or the like, unit dosecontainers, blister packs, and strip packs.

An active ingredient can be combined in an intimate admixture with apharmaceutical carrier according to various pharmaceutical compoundingtechniques. The carrier can take a wide variety of forms depending onthe form of preparation desired for administration. In preparing thecompositions for an oral dosage form, any of the usual pharmaceuticalmedia can be employed as carriers, such as, for example, water, glycols,oils, alcohols, flavoring agents, preservatives, coloring agents, andthe like in the case of oral liquid preparations (such as suspensions,solutions, and elixirs) or aerosols; or carriers such as starches,sugars, micro-crystalline cellulose, diluents, granulating agents,lubricants, binders, and disintegrating agents can be used in the caseof oral solid preparations, in some embodiments without employing theuse of lactose. For example, suitable carriers include powders,capsules, and tablets, with the solid oral preparations. If desired,tablets can be coated by standard aqueous or nonaqueous techniques.

Binders suitable for use in pharmaceutical compositions and dosage formsmay include, but are not limited to, corn starch, potato starch, orother starches, gelatin, natural and synthetic gums such as acacia,sodium alginate, alginic acid, other alginates, powdered tragacanth,guar gum, cellulose and its derivatives (e.g., ethyl cellulose,cellulose acetate, carboxymethyl cellulose calcium, sodium carboxymethylcellulose), polyvinyl pyrrolidone, methyl cellulose, pre-gelatinizedstarch, hydroxypropyl methyl cellulose, microcrystalline cellulose, andmixtures thereof.

Examples of suitable fillers for use in the pharmaceutical compositionsand dosage forms disclosed herein may include, but are not limited to,talc, calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.

Disintegrants may be used in the compositions of the disclosure toprovide tablets that may disintegrate when exposed to an aqueousenvironment. Too much of a disintegrant may produce tablets which maydisintegrate in the bottle. Too little may be insufficient fordisintegration to occur and may thus alter the rate and extent ofrelease of the active ingredient(s) from the dosage form. Thus, acertain amount of disintegrant that is neither too little nor too muchto detrimentally alter the release of the active ingredient(s) may beused to form the dosage forms of the glycopolymers disclosed herein. Theamount of disintegrant used may vary based upon the type of formulationand mode of administration. About 0.5 to about 15 weight percent ofdisintegrant, or about 1 to about 5 weight percent of disintegrant, maybe used in the pharmaceutical composition. Disintegrants that can beused to form pharmaceutical compositions and dosage forms of thedisclosure may include, but are not limited to, agar-agar, alginic acid,calcium carbonate, microcrystalline cellulose, croscarmellose sodium,crospovidone, polacrilin potassium, sodium starch glycolate, potato ortapioca starch, other starches, pre-gelatinized starch, other starches,clays, other algins, other celluloses, gums or mixtures thereof.

Lubricants may be used to form pharmaceutical compositions and dosageforms of the disclosure. Non-limiting examples of lubricants mayinclude, but not limited to, calcium stearate, magnesium stearate,mineral oil, light mineral oil, glycerin, sorbitol, mannitol,polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate,talc, hydrogenated vegetable oil (e.g., peanut oil, cottonseed oil,sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zincstearate, ethyl oleate, ethyl laureate, agar, or mixtures thereof.Additional lubricants may include, for example, a syloid silica gel, acoagulated aerosol of synthetic silica, or mixtures thereof. A lubricantcan optionally be added, in an amount of less than about 1 weightpercent of the pharmaceutical composition.

When aqueous suspensions and/or elixirs are desired for oraladministration, the active ingredient therein may be combined withvarious sweetening or flavoring agents, coloring matter or dyes and, ifso desired, emulsifying and/or suspending agents, together with suchdiluents as water, ethanol, propylene glycol, glycerin and variouscombinations thereof.

The tablets can be uncoated or coated by a number of techniques to delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearate canbe employed. Formulations for oral use can also be presented as hardgelatin capsules wherein the active ingredient is mixed with an inertsolid diluent, for example, calcium carbonate, calcium phosphate orkaolin, or as soft gelatin capsules wherein the active ingredient ismixed with water or an oil medium, for example, peanut oil, liquidparaffin or olive oil.

Surfactant can be used to form pharmaceutical compositions and dosageforms of the disclosure. Examples of surfactants may include, but notlimited to, hydrophilic surfactants, lipophilic surfactants, andmixtures thereof. That is, a mixture of hydrophilic surfactants may beemployed, a mixture of lipophilic surfactants may be employed, or amixture of at least one hydrophilic surfactant and at least onelipophilic surfactant may be employed.

An empirical parameter used to characterize the relative hydrophilicityand hydrophobicity of non-ionic amphiphilic compounds is thehydrophilic-lipophilic balance (“HLB” value). A suitable hydrophilicsurfactant may generally have an HLB value of at least about 10, whilesuitable lipophilic surfactants may generally have an HLB value of orless than about 10. Surfactants with lower HLB values are morelipophilic or hydrophobic, and have greater solubility in oils, whilesurfactants with higher HLB values are more hydrophilic, and havegreater solubility in aqueous solutions. Hydrophilic surfactants aregenerally considered to be those compounds having an HLB value greaterthan about 10, as well as anionic, cationic, or zwitterionic compoundsfor which the HLB scale is not generally applicable. Similarly,lipophilic (i.e., hydrophobic) surfactants are compounds having an HLBvalue equal to or less than about 10. However, HLB value of a surfactantis merely a rough guide generally used to enable formulation ofindustrial, pharmaceutical and cosmetic emulsions.

Hydrophilic surfactants may be either ionic or non-ionic. Ionicsurfactants may include, but not limited to, alkylammonium salts,fusidic acid salts, fatty acid derivatives of amino acids,oligopeptides, and polypeptides, glyceride derivatives of amino acids,oligopeptides, and polypeptides, lecithins and hydrogenated lecithins,lysolecithins and hydrogenated lysolecithins, phospholipids andderivatives thereof, lysophospholipids and derivatives thereof,carnitine fatty acid ester salts, salts of alkylsulfates, fatty acidsalts, sodium docusate, acyl lactylates, mono- and di-acetylatedtartaric acid esters of mono- and di-glycerides, succinylated mono- anddi-glycerides, citric acid esters of mono- and di-glycerides; andmixtures thereof.

Within the aforementioned group, ionic surfactants may include, by wayof example: lecithins, lysolecithin, phospholipids, lysophospholipidsand derivatives thereof; carnitine fatty acid ester salts; salts ofalkylsulfates; fatty acid salts; sodium docusate; acylactylates; mono-and di-acetylated tartaric acid esters of mono- and di-glycerides;succinylated mono- and di-glycerides; citric acid esters of mono- anddi-glycerides; and mixtures thereof.

Ionic surfactants may be the ionized forms of lecithin, lysolecithin,phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol,phosphatidic acid, phosphatidylserine, lysophosphatidylcholine,lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidicacid, lysophosphatidylserine, PEG-phosphatidylethanolamine,PVP-phosphatidylethanolamine, lactylic esters of fatty acids,stearoyl-2-lactylate, stearoyl lactylate, succinylated monoglycerides,mono/diacetylated tartaric acid esters of mono/diglycerides, citric acidesters of mono/diglycerides, cholylsarcosine, caproate, caprylate,caprate, laurate, myristate, palmitate, oleate, ricinoleate, linoleate,linolenate, stearate, lauryl sulfate, teracecyl sulfate, docusate,lauroyl carnitines, palmitoyl carnitines, myristoyl carnitines, andsalts and mixtures thereof.

Hydrophilic non-ionic surfactants may include, but not limited to,alkylglucosides; alkylmaltosides; alkylthioglucosides; laurylmacrogolglycerides; polyoxyalkylene alkyl ethers such as polyethyleneglycol alkyl ethers; polyoxyalkylene alkylphenols such as polyethyleneglycol alkyl phenols; polyoxyalkylene alkyl phenol fatty acid esterssuch as polyethylene glycol fatty acids monoesters and polyethyleneglycol fatty acids diesters; polyethylene glycol glycerol fatty acidesters; polyglycerol fatty acid esters; polyoxyalkylene sorbitan fattyacid esters such as polyethylene glycol sorbitan fatty acid esters;hydrophilic transesterification products of a polyol with at least onemember of the group consisting of glycerides, vegetable oils,hydrogenated vegetable oils, fatty acids, and sterols; polyoxyethylenesterols, derivatives, and analogues thereof; polyoxyethylated vitaminsand derivatives thereof; polyoxyethylene-polyoxypropylene blockcopolymers; and mixtures thereof; polyethylene glycol sorbitan fattyacid esters and hydrophilic transesterification products of a polyolwith at least one member of the group consisting of triglycerides,vegetable oils, and hydrogenated vegetable oils. The polyol may beglycerol, ethylene glycol, polyethylene glycol, sorbitol, propyleneglycol, pentaerythritol, or a saccharide.

Other hydrophilic-non-ionic surfactants may include, without limitation,PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate,PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32distearate, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25glyceryl trioleate, PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30glyceryl laurate, PEG-20 glyceryl stearate, PEG-20 glyceryl oleate,PEG-30 glyceryl oleate, PEG-30 glyceryl laurate, PEG-40 glyceryllaurate, PEG-40 palm kernel oil, PEG-50 hydrogenated castor oil, PEG-40castor oil, PEG-35 castor oil, PEG-60 castor oil, PEG-40 hydrogenatedcastor oil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG-6caprate/caprylate glycerides, PEG-8 caprate/caprylate glycerides,polyglyceryl-10 laurate, PEG-30 cholesterol, PEG-25 phyto sterol, PEG-30soya sterol, PEG-20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitanlaurate, polysorbate 20, polysorbate 80, POE-9 lauryl ether, POE-23lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearylether, tocopheryl PEG-100 succinate, PEG-24 cholesterol,polyglyceryl-10oleate, Tween 40, Tween 60, sucrose monostearate, sucrosemonolaurate, sucrose monopalmitate, PEG 10-100 nonyl phenol series, PEG15-100 octyl phenol series, and poloxamers.

Lipophilic surfactants may include, by way of example only: fattyalcohols; glycerol fatty acid esters; acetylated glycerol fatty acidesters; lower alcohol fatty acids esters; propylene glycol fatty acidesters; sorbitan fatty acid esters; polyethylene glycol sorbitan fattyacid esters; sterols and sterol derivatives; polyoxyethylated sterolsand sterol derivatives; polyethylene glycol alkyl ethers; sugar esters;sugar ethers; lactic acid derivatives of mono- and di-glycerides;hydrophobic transesterification products of a polyol with at least onemember of the group consisting of glycerides, vegetable oils,hydrogenated vegetable oils, fatty acids and sterols; oil-solublevitamins/vitamin derivatives; and mixtures thereof. Within this group,preferred lipophilic surfactants include glycerol fatty acid esters,propylene glycol fatty acid esters, and mixtures thereof, or arehydrophobic transesterification products of a polyol with at least onemember of the group consisting of vegetable oils, hydrogenated vegetableoils, and triglycerides.

In one embodiment, the composition may include a solubilizer to ensuregood solubilization and/or dissolution of the compound of the presentdisclosure and to minimize the precipitation of the compound. Asolubilizer may also be added to increase the solubility of thehydrophilic drug and/or other components, such as surfactants, or tomaintain the composition as a stable or homogeneous solution ordispersion.

Examples of solubilizers may include, but are not limited to alcoholsand polyols, such as ethanol, isopropanol, butanol, benzyl alcohol,ethylene glycol, propylene glycol, butanediols and isomers thereof,glycerol, pentaerythritol, sorbitol, mannitol, transcutol, dimethylisosorbide, polyethylene glycol, polypropylene glycol, polyvinylalcohol,hydroxypropyl methylcellulose and other cellulose derivatives,cyclodextrins and cyclodextrin derivatives; ethers of polyethyleneglycols having an average molecular weight of about 200 to about 6000,such as tetrahydrofurfuryl alcohol PEG ether (glycofurol) or methoxyPEG; amides and other nitrogen-containing compounds such as2-pyrrolidone, 2-piperidone, ε-caprolactam, N-alkylpyrrolidone,N-hydroxyalkylpyrrolidone, N-alkylpiperidone, N-alkylcaprolactam,dimethylacetamide and polyvinylpyrrolidone; esters such as ethylpropionate, tributylcitrate, acetyl triethylcitrate, acetyl tributylcitrate, triethylcitrate, ethyl oleate, ethyl caprylate, ethyl butyrate,triacetin, propylene glycol monoacetate, propylene glycol diacetate,ε-caprolactone and isomers thereof, δ-valerolactone and isomers thereof,β-butyrolactone and isomers thereof; and other solubilizers known in theart, such as dimethyl acetamide, dimethyl isosorbide, N-methylpyrrolidones, monooctanoin, diethylene glycol monoethyl ether, andwater.

Mixtures of solubilizers may also be used. Examples may include, but notlimited to, triacetin, triethylcitrate, ethyl oleate, ethyl caprylate,dimethylacetamide, N-methylpyrrolidone, N-hydroxyethylpyrrolidone,polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxypropylcyclodextrins, ethanol, polyethylene glycol 200-100, glycofurol,transcutol, propylene glycol, and dimethyl isosorbide. Particularlypreferred solubilizers include sorbitol, glycerol, triacetin, ethylalcohol, PEG-400, glycofurol and propylene glycol.

The amount of solubilizer that can be included may vary. In some cases,the amount of a given solubilizer may be limited to a bioacceptableamount. In some cases, it may be advantageous to include amounts ofsolubilizers far in excess of bioacceptable amounts, for example tomaximize the concentration of the drug, with excess solubilizer removedprior to providing the composition to a subject using conventionaltechniques, such as distillation or evaporation. Thus, if present, thesolubilizer can be in a weight ratio of 10%, 25%, 50%, 100%, or up toabout 200% by weight, based on the combined weight of the drug, andother excipients. If desired, very small amounts of solubilizer may alsobe used, such as 5%, 2%, 1% or even less. Typically, the solubilizer maybe present in an amount of about 1% to about 100%, more typically about5% to about 25% by weight.

The composition can further include one or more pharmaceuticallyacceptable additives and excipients. Such additives and excipients mayinclude, without limitation, detackifiers, anti-foaming agents,buffering agents, polymers, antioxidants, preservatives, chelatingagents, viscomodulators, tonicifiers, flavorants, colorants, odorants,opacifiers, suspending agents, binders, fillers, plasticizers,lubricants, and mixtures thereof.

In addition, an acid or a base may be incorporated into the compositionto facilitate processing, to enhance stability, or for other reasons.Examples of pharmaceutically acceptable bases may include amino acids,amino acid esters, ammonium hydroxide, potassium hydroxide, sodiumhydroxide, sodium hydrogen carbonate, aluminum hydroxide, calciumcarbonate, magnesium hydroxide, magnesium aluminum silicate, syntheticaluminum silicate, synthetic hydrocalcite, magnesium aluminum hydroxide,diisopropylethylamine, ethanolamine, ethylenediamine, triethanolamine,triethylamine, triisopropanolamine, trimethylamine, tris(hydroxymethyl)aminomethane (TRIS) and the like. Additional examples of bases may besalts of a pharmaceutically acceptable acid, such as acetic acid,acrylic acid, adipic acid, alginic acid, alkanesulfonic acid, aminoacids, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonicacid, citric acid, fatty acids, formic acid, fumaric acid, gluconicacid, hydroquinosulfonic acid, isoascorbic acid, lactic acid, maleicacid, oxalic acid, para-bromophenylsulfonic acid, propionic acid,p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid,tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid,uric acid, and the like. Salts of polyprotic acids, such as sodiumphosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphatecan also be used. When the base is a salt, the cation can be anyconvenient and pharmaceutically acceptable cation, such as ammonium,alkali metals, alkaline earth metals, and the like. Example may include,but not limited to, sodium, potassium, lithium, magnesium, calcium andammonium.

Acids may also be used in pharmaceutical compositions, such aspharmaceutically acceptable organic or inorganic acids. Examples ofinorganic acids may include hydrochloric acid, hydrobromic acid,hydriodic acid, sulfuric acid, nitric acid, boric acid, phosphoric acid,and the like. Examples of suitable organic acids include acetic acid,acrylic acid, adipic acid, alginic acid, alkanesulfonic acids, aminoacids, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonicacid, citric acid, fatty acids, formic acid, fumaric acid, gluconicacid, hydroquinosulfonic acid, isoascorbic acid, lactic acid, maleicacid, methanesulfonic acid, oxalic acid, para-bromophenylsulfonic acid,propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid,succinic acid, tannic acid, tartaric acid, thioglycolic acid,toluenesulfonic acid, uric acid and the like.B. Pharmaceutical Compositions for Injection

The present invention also provides a pharmaceutical composition forinjection comprising a compound of the disclosure and a pharmaceuticalexcipient suitable for injection. Components and amounts of agents inthe compositions are as described herein.

The forms in which the pharmaceutical compositions of the presentdisclosure may be incorporated for administration by injection includeaqueous or oil suspensions, or emulsions, with sesame oil, corn oil,cottonseed oil, or peanut oil, as well as elixirs, mannitol, dextrose,or a sterile aqueous solution, and similar pharmaceutical vehicles.

Aqueous solutions in saline may be used for injection. Ethanol,glycerol, propylene glycol, liquid polyethylene glycol, and the like(and suitable mixtures thereof), cyclodextrin derivatives, and vegetableoils may also be employed. The proper fluidity can be maintained, forexample, by the use of a coating, such as lecithin, for the maintenanceof the required particle size in the case of dispersion and by the useof surfactants. The prevention of the action of microorganisms can bebrought about by various antibacterial and antifungal agents, forexample, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, andthe like.

Sterile injectable solutions may be prepared by incorporating thecompound of the present invention in the required amount in theappropriate solvent with various other ingredients as enumerated above,as required, followed by filtered sterilization. Generally, dispersionsmay be prepared by incorporating the various sterilized activeingredients into a sterile vehicle which contains the basic dispersionmedium and the required other ingredients from those enumerated above.In the case of sterile powders for the preparation of sterile injectablesolutions, certain methods of preparation may be vacuum-drying andfreeze-drying techniques which yield a powder of the active ingredientplus any additional desired ingredient from a previouslysterile-filtered solution thereof.

C. Pharmaceutical Compositions for Topical (e.g., Transdermal) Delivery

In some embodiments, the disclosure provides a pharmaceuticalcomposition for transdermal delivery containing a compound of thepresent disclosure and a pharmaceutical excipient for transdermaldelivery.

Compositions of the present disclosure can be formulated intopreparations in solid, semi-solid, or liquid forms suitable for local ortopical administration, such as gels, water soluble jellies, creams,lotions, suspensions, foams, powders, slurries, ointments, solutions,oils, pastes, suppositories, sprays, emulsions, saline solutions,dimethylsulfoxide (DMSO)-based solutions. In general, carriers withhigher densities are capable of providing an area with a prolongedexposure to the active ingredients. In contrast, a solution formulationmay provide more immediate exposure of the active ingredient to thechosen area.

The pharmaceutical compositions may also comprise suitable solid or gelphase carriers or excipients, which are compounds that allow increasedpenetration of, or assist in the delivery of, therapeutic moleculesacross the stratum corneum permeability barrier of the skin. Examples ofsuch carriers and excipients may include, but not limited to, humectants(e.g., urea), glycols (e.g., propylene glycol), alcohols (e.g.,ethanol), fatty acids (e.g., oleic acid), surfactants (e.g., isopropylmyristate and sodium lauryl sulfate), pyrrolidones, glycerolmonolaurate, sulfoxides, terpenes (e.g., menthol), amines, amides,alkanes, alkanols, water, calcium carbonate, calcium phosphate, varioussugars, starches, cellulose derivatives, gelatin, and polymers such aspolyethylene glycols.

Another exemplary formulation for use in the methods of the presentdisclosure employs transdermal delivery devices (“patches”). Suchtransdermal patches may be used to provide continuous or discontinuousinfusion of a compound of the present invention in controlled amounts,either with or without another agent.

D. Pharmaceutical Compositions for Inhalation

Compositions for inhalation or insufflation may include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically acceptable excipients as describedelsewhere herein. The compositions may be administered by the oral ornasal respiratory route for local or systemic effect. Compositions inpharmaceutically acceptable solvents may be nebulized by use of inertgases. Nebulized solutions may be inhaled directly from the nebulizingdevice or the nebulizing device may be attached to a face mask tent, orintermittent positive pressure breathing machine. Solution, suspension,or powder compositions may be administered, preferably orally ornasally, from devices that deliver the formulation in an appropriatemanner.

E. Pharmaceutical Compositions for Other Administration Methods

Pharmaceutical compositions may also be prepared from compositionsdescribed herein and one or more pharmaceutically acceptable excipientssuitable for sublingual, buccal, rectal, intraosseous, intraocular,intranasal, epidural, or intraspinal administration. A number of methodsmay be used to prepare for such pharmaceutical compositions. See, e.g.,See, e.g., Anderson, Philip O.; Knoben, James E.; Troutman, William G,eds., Handbook of Clinical Drug Data, Tenth Edition, McGraw-Hill, 2002;Pratt and Taylor, eds., Principles of Drug Action, Third Edition,Churchill Livingston, N.Y., 1990; Katzung, ed., Basic and ClinicalPharmacology, Ninth Edition, McGraw Hill, 20037ybg; Goodman and Gilman,eds., The Pharmacological Basis of Therapeutics, Tenth Edition, McGrawHill, 2001; Remingtons Pharmaceutical Sciences, 20th Ed., LippincottWilliams & Wilkins, 2000; Martindale, The Extra Pharmacopoeia,Thirty-Second Edition (The Pharmaceutical Press, London, 1999); all ofwhich are incorporated by reference herein in their entirety.

Administration of the compounds or pharmaceutical composition of thepresent disclosure can be effected by any method that enables deliveryof the compounds to the site of action. These methods may include oralroutes, intraduodenal routes, parenteral injection (includingintravenous, intraarterial, subcutaneous, intramuscular, intravascular,intraperitoneal or infusion), topical (e.g. transdermal application),rectal administration, via local delivery by catheter or stent orthrough inhalation. Compounds can also be administered intraadiposallyor intrathecally.

As described herein, the amount of the compound administered may bedependent upon, the subject being treated, the severity of the disorderor condition, the rate of administration, the disposition of thecompound and the discretion of the prescribing physician. However, aneffective dosage is in the range of about 0.001 to about 100 mg per kgbody weight per day, preferably about 1 to about 35 mg/kg/day, in singleor divided doses. For a 70 kg human, this would amount to about 0.05 to7 g/day, preferably about 0.05 to about 2.5 g/day. In some instances,dosage levels below the lower limit of the aforementioned range may bemore than adequate, while in other cases still larger doses may beemployed without causing any harmful side effect, e.g. by dividing suchlarger doses into several small doses for administration throughout theday.

In some embodiments, a pharmaceutical composition may be administered ina single dose. In many cases, such administration may be by injection,e.g., intravenous injection, in order to introduce the agent quickly.However, other routes may be used as appropriate. A single dose of thepharmaceutical composition may also be used for treatment of an acutecondition.

In some embodiments, a pharmaceutical composition may be administered inmultiple doses. Dosing may be about once, twice, three times, fourtimes, five times, six times, or more than six times per day. Dosing maybe about once a month, once every two weeks, once a week, or once everyother day. In some embodiments a composition and another agent may beadministered together about once per day to about 6 times per day. Insome embodiments the administration of a composition and an agentcontinues for less than about 7 days. In some embodiments theadministration may continue for more than about 6, 10, 14, 28 days, twomonths, six months, or one year. In some embodiments, continuous dosingmay be achieved and maintained as long as necessary.

Administration of the composition may continue as long as necessary. Insome embodiments, a composition may be administered for more than 1, 2,3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, a composition may beadministered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In someembodiments, a composition may be administered chronically on an ongoingbasis, e.g., for the treatment of chronic effects.

An effective amount of a compound may be administered in either singleor multiple doses by any of the accepted modes of administration ofagents having similar utilities, including rectal, buccal, intranasaland transdermal routes, by intra-arterial injection, intravenously,intraperitoneally, parenterally, intramuscularly, subcutaneously,orally, topically, or as an inhalant.

The compositions of the disclosure may also be delivered via animpregnated or coated device such as a stent, or an artery-insertedcylindrical polymer. Such a method of administration may, for example,aid in the prevention or amelioration of restenosis following proceduressuch as balloon angioplasty. Without being bound by theory, acomposition may slow or inhibit the migration and proliferation ofsmooth muscle cells in the arterial wall which contribute to restenosis.A composition may be administered, for example, by local delivery fromthe struts of a stent, from a stent graft, from grafts, or from thecover or sheath of a stent. In some embodiments, a composition may beadmixed with a matrix. Such a matrix may be a polymeric matrix, and mayserve to bond the compound to the stent. Polymeric matrices may include,for example, lactone-based polyesters or copolyesters such aspolylactide, polycaprolactonglycolide, polyorthoesters, polyanhydrides,polyaminoacids, polysaccharides, polyphosphazenes, poly (ether-ester)copolymers (e.g. PEO-PLLA); polydimethylsiloxane,poly(ethylene-vinylacetate), acrylate-based polymers or copolymers (e.g.polyhydroxyethyl methylmethacrylate, polyvinyl pyrrolidinone),fluorinated polymers such as polytetrafluoroethylene and celluloseesters. Matrices may be degradable or nondegradable, releasing theingredients. Glycopolymers may be applied to the surface of the stent byvarious methods such as dip/spin coating, spray coating, dip-coating,and/or brush-coating. The glycopolymers may be applied in a solvent andthe solvent may be allowed to evaporate, thus forming a layer ofcompound onto the stent. Alternatively, the glycopolymers may be locatedin the body of the stent or graft, for example in microchannels ormicropores. When implanted, the glycopolymer diffuses out of the body ofthe stent to contact the arterial wall. Such stents may be prepared bydipping a stent manufactured to contain such micropores or microchannelsinto a solution of the compound in a suitable solvent, followed byevaporation of the solvent. Excess drug on the surface of the stent maybe removed via an additional brief solvent wash. In some embodiments,compound may be covalently linked to a stent or graft. A covalent linkermay be used which may degrade in vivo, leading to the release of thecompound of the invention. Any bio-labile linkage may be used for such apurpose, such as ester, amide or anhydride linkages. Compounds mayadditionally be administered intravascularly from a balloon used duringangioplasty. Extravascular administration of the compounds via thepericard or via advential application of formulations of the disclosuremay also be performed to decrease restenosis.

A variety of stent devices which may be used as described are disclosed,for example, in the following references, all of which are herebyincorporated by reference: U.S. Pat. No. 5,451,233; U.S. Pat. No.5,040,548; U.S. Pat. No. 5,061,273; U.S. Pat. No. 5,496,346; U.S. Pat.No. 5,292,331; U.S. Pat. No. 5,674,278; U.S. Pat. No. 3,657,744; U.S.Pat. No. 4,739,762; U.S. Pat. No. 5,195,984; U.S. Pat. No. 5,292,331;U.S. Pat. No. 5,674,278; U.S. Pat. No. 5,879,382; U.S. Pat. No.6,344,053.

Kits

The present disclosure also provides kits. The kits include one or morecompounds of the disclosure as described elsewhere herein, in packaging,and written material that can include instructions for use, discussionof clinical studies, listing of side effects, and the like. Such kitsmay also include information, such as scientific literature references,package insert materials, clinical trial results, and/or summaries ofthese and the like, which indicate or establish the activities and/oradvantages of the composition, and/or which describe dosing,administration, side effects, drug interactions, or other informationuseful to the health care provider. Such information may be based on theresults of various studies, for example, studies using experimentalanimals involving in vivo models and studies based on human clinicaltrials. The kit may further contain another agent. In some embodiments,the compound of the present invention and the agent are provided asseparate compositions in separate containers within the kit. In someembodiments, the compound of the present invention and the agent areprovided as a single composition within a container in the kit. Suitablepackaging and additional articles for use (e.g., measuring cup forliquid preparations, foil wrapping to minimize exposure to air, and thelike) are known in the art and may be included in the kit. Kitsdescribed herein can be provided, marketed and/or promoted to healthproviders, including physicians, nurses, pharmacists, formularyofficials, and the like. Kits may also, in some embodiments, be marketeddirectly to the consumer.

Substrates

The present disclosure also contemplates many uses for glypolymersattached to, associated with, or immobilized on one or more substrates.In some embodiments, the substrate may include a solid support, such asa membrane, filter, microscope slide, microwell, array, tube, bead, beadarray, or the like. The substrate may be made of various materials,including, but not limited to paper, cellulose, nylon, polystyrene,polycarbonate, plastics, glass, ceramic, stainless steel, or the like.In some cases, the substrate may have a rigid or semi-rigid surface. Insome cases, the substrate may be spherical (e.g., bead) or substantiallyplanar (e.g., flat surface) with appropriate wells, raised regions,etched trenches, or the like. In some cases, the substrate may include agel or matrix in which glycopolymers may be embedded.

The substrate may be a part of, or in the form of a microparticle,nanoparticle, bandage, suture, catheter, stent, valve, pacemaker,implantable defibrillator, conduit, cannula, appliance, scaffold,central line (which may be a peripherally inserted central catheter(PICC or PIC line)), pessary, tube, drain, shunt, trochar, plug, orother implant or medical or surgical device. In some cases, the cathetermay be a pulmonary artery, pericardial, pleural, urinary orintra-abdominal catheter. In some cases, the drain may be acerebrospinal fluid drain. In some cases, the tube may be atracheostomy, endotracheal or chest tube. In some other embodiments, thesubstrate may be a part of, or in the form of an implant, a rod (e.g. aspinal rod such as a posterior spinal rod), a plate, a screw, washer,wire, pin, internal fixation devices (e.g. fracture fixation devices),or other implantable orthopedic hardware known in the art.

Also provided in the present disclosure is that substrates may beflexible such that they may readily conform or bend to adopt a desiredshape or configuration under conditions of use, or they may be rigidsuch that significant force may be required to cause an alteration inshape. In some embodiments the substrate may maintain its shape whensupported at only one point or end. The surface could be substantiallysmooth or could be rough and/or comprise crevices.

EXAMPLES Example 1: Synthesis and Characterization of Compounds

Exemplary methods of synthesizing the disaccharide compounds 5 and 15,and intermediates thereof, is shown in FIG. 2.

Methyl 3-O-benzyl-L-idopyranosyluronate (9)

Compound 9 was prepared in six steps from the commercially availablediacetone glucose (Sigma Aldrich) using a number of procedures. (Seee.g., Orgueira, H. A. et al., Chem. Eur. J. 2003, 9, 140; Lohman, G. J.S. et al., J. Org. Chem. 2003, 68, 7559). The analytical data were inagreement with the reported spectra.

Methyl 1,2,4-tri-O-acetyl-3-O-benzyl-β-L-idopyranosyluronate (10)

Compound 9 (0.30 g, 1.0 mmol) was added to CH₂Cl₂ (5.5 mL) at 0° C., andthe solution was cooled to −40° C. 4-Dimethylaminopyridine (120 mg, 0.10mmol) was added, followed by pyridine (700 μL, 10 mmol). Acetyl chloride(470 μL, 6.0 mmol) was then added dropwise to the reaction mixture,which was stirred for 10 h at −40° C. The reaction was quenched withaqueous NaHCO₃ (50 mL), extracted with CH₂Cl₂ (2.0×50 mL), andsubsequently washed with H₂O, 1M H₂SO₄, and then H₂O (50 mL for eachwash). The combined organic layers were dried over MgSO₄ andconcentrated under reduced pressure. Purification by silica gel flashchromatography (3:1 hexanes:EtOAc) afforded compound 10 (0.40 g) inquantitative yield. The analytical data were in agreement withpreviously reported spectra.³ ESI-TOF HRMS: m/z calcd for C₂₀H₂₃O₁₀[M+H]—H₂ 423.1286. found: 423.1286.

Methyl 4-O-acetyl-3-O-benzyl-β-L-idopyranuronate1,2-(methyl-orthoacetate) (11)

TiBr₄ (8.1 g, 22 mmol) was added to a solution of compound 10 (6.9 g, 16mmol) in CH₂Cl₂ (360 mL), and the reaction was stirred for 16 h atambient temperature with exclusion of light. The reaction was quenchedwith ice-cold H₂O (2.0×500 mL), filtered through Celite, andconcentrated under reduced pressure. The resulting brown oil wasimmediately used in the next reaction without further purification. Thecrude bromide intermediate (16 mmol) was dissolved in CH₂Cl₂ (220 mL).2,4,6-Collidine (11 mL, 80 mmol) and methanol (8.0 mL) were added tothis solution, and the reaction was stirred for 14 h at room temperature(rt). The reaction mixture was then diluted with CH₂Cl₂ (500 mL), washedwith aqueous NaHCO₃ and H₂O (200 mL each), dried over MgSO₄, andconcentrated under reduced pressure. Purification by silica gel flashchromatography (6:1 hexanes:EtOAc+1% Et₃N) afforded 11 (7.6 g, 75% over2 steps) as a light yellow oil. ¹H NMR (500 MHz; CDCl₃): δ 7.44-7.30 (m,5H, OCH₂Ph), 5.57 (d, J=2.7 Hz, 1H, H-1), 5.20 (dt, J=2.7, 1.3 Hz, 1H,H-4), 4.82 (d, J=11.7 Hz, 1H, OCH₂Ph), 4.69 (d, J=11.7 Hz, 1H, OCH₂Ph),4.56 (d, J=1.4 Hz, 1H, H-5), 4.15 (dd, J=2.7, 1.9 Hz, 1H, H-3), 4.09(ddd, J=2.9, 1.9, 1.2 Hz, 1H, H-2), 3.79 (s, 3H, CO₂CH₃), 3.26 (s, 3H,OCH₃), 2.05 (s, 3H, OCOCH₃), 1.74 (s, 3H, CH₃); ¹³C NMR (125 MHz;CDCl₃): δ 170.1, 168.1, 136.8, 128.6, 128.4, 128.0, 96.6, 77.3, 76.1,72.9, 71.3, 69.6, 68.9, 52.6, 49.1, 25.0, 20.1; ESI-TOF HRMS: m/z calcdfor C₁₉H₂₃O₉ [M+H]—H₂ 395.1342. found: 395.1354.

Methyl 3-O-benzyl-β-L-idopyranuronate 1,2-(methyl-orthoacetate) (12)

Compound 11 (7.2 g, 18 mmol) was dissolved in methanol (90 mL) andcooled to −10° C. A 0.5 M solution NaOMe (1.8 mL, 0.91 mmol) was added,and the reaction mixture was stirred at −10° C. for 2 h and at 5° C.overnight. The solution was diluted with CH₂Cl₂ (200 mL) at 5° C.,quenched with aqueous NaHCO₃ and H₂O (500 mL each), and then extractedwith (3.0×250 mL). The organic fractions were dried over MgSO₄ andconcentrated under reduced pressure. Purification by silica gel flashchromatography (4:1→1:1 hexanes:EtOAc+1% Et₃N) yielded 12 (9.5 g, 80%)as a clear oil. ¹H NMR (500 MHz; CDCl₃): δ 7.36-7.34 (m, 5H, OCH₂Ph),5.51 (d, J=2.4 Hz, 1H, H-1), 4.72 (d, J=11.7 Hz, 1H, OCH₂Ph), 4.62 (d,J=11.7 Hz, 1H, OCH₂Ph), 4.52 (s, 1H, H-4), 4.15-4.08 (m, 3H, H-2, H-5),3.81 (s, 3H, CO₂CH₃), 3.30 (s, 3H, OCH₃), 2.78 (d, J=11.4 Hz, 1H, H-3),1.76 (s, 3H, CH₃); ¹³C NMR (125 MHz; CDCl₃): δ 168.3, 136.8, 128.7,128.4, 127.9, 96.8, 75.8, 73.0, 72.9, 71.8, 67.0, 52.5, 50.3, 24.4;ESI-TOF HRMS: m/z calcd for C₁₇H₂₁O₈ [M+H]—H₂ 353.1236. found: 353.1226.

Methyl 3-O-benzyl-4-O-tert-butyldimethylsilyl-β-L-idopyranuronate1,2-(methyl-orthoace-tate) (13)

Compound 12 (230 mg, 0.64 mmol) was dissolved in pyridine (7.8 mL) andthe solution was cooled to −10° C. TBSOTf (1.5 mL, 0.65 mmol) was added,and the reaction was stirred overnight at 0° C. The reaction was dilutedwith CH₂Cl₂ (100 mL), quenched with aqueous NaHCO₃ (100 mL), andextracted with EtOAc (3.0×50 mL). The combined organic fractions weredried over MgSO₄ and concentrated under reduced pressure. Purificationby silica gel flash chromatography (7:1 hexanes:EtOAc+1% Et₃N) yieldedcompound 13 (380 mg, 92%) as a clear oil. ¹H NMR (500 MHz; CDCl₃): δ7.47 (m, 5H, OCH₂Ph), 5.62 (d, J=2.5 Hz, 1H, H-1), 4.80 (d, J=12 Hz, 1H,OCH₂Ph), 4.75 (d, J=12 Hz, 1H, OCH₂Ph), 4.51 (s, 1H, H-4), 4.21 (s, 1H,H-5), 4.20 (d, J=1 Hz, 1H, H-4), 3.98 (s, 1H, H-3), 3.89 (s, 3H,CO₂CH₃), 3.40 (s, 3H, OCH₃), 1.84 (s, 3H, CH₃), 0.94 (s, 9H, SiC(CH₃)₃),0.08 (s, 3H, SiCH₃), 0.06 (s, 3H, SiCH₃); ¹³C NMR (125 MHz; CDCl₃): δ169.6, 137.0, 128.9, 128.6, 128.2, 124.6, 97.1, 76.3. 74.6, 72.8, 72.5,67.9, 52.4, 49.5, 29.9, 25.7, 25.6, −4.4, −5.2; TOF HRMS ES m/z calcdfor C₂₃H₃₆O₈SiNa [M+Na]⁺: 491.2077. found: 491.2070.

Methyl(dibutylphosphate-2-O-acetyl-3-O-benzyl-4-O-tert-butyldimethylsilyl-α-L-idopyra-nosid)uronate(6)

Compound 13 (140 mg, 0.29 mmol) was dissolved in CH₂Cl₂ (7.3 mL) at rt.Freshly activated 4 Å molecular sieves (290 mg) were added, and thesolution was stirred for 15 min. Dibutylphosphate (0.54 mL, 2.9 mmol)was added slowly, and the reaction mixture was stirred overnight. Afterconfirming that the reaction was complete by TLC, the reaction wasquenched with triethylamine (2.0 mL) and concentrated under reducedpressure. Silica gel flash chromatography (5:1→3:1 hexanes:EtOAc+1%Et₃N) afforded the desired product (170 mg) in quantitative yield. ¹HNMR (500 MHz; CDCl₃): δ 7.36-7.35 (m, 5H, OCH₂Ph), 5.82 (d, J=7.2 Hz,1H, H-1), 4.97 (m, 1H, H-2), 4.86 (d, J=2.7 Hz, 1H, H-5), 4.78 (d, J=12Hz, 1H, OCH₂Ph), 4.62 (d, J=12 Hz, 1H, OCH₂Ph), 4.09-3.99 (m, 5H, H-4,P(OCH₂CH₂CH₃)₂), 3.77 (s, 3H, CO₂CH₃), 3.62 (m, 1H, H-3), 2.04 (s, 3H,COCH₃), 1.64-1.60 (m, 4H, P(OCH₂CH₂CH₂CH₃)₂), 1.40-1.25 (m, 4H,P(OCH₂CH₂CH₂CH₃)₂), 0.96-0.88 (m, 6H, P(OCH₂CH₂CH₂CH₃)₂), 0.81 (s, 9H,SiC(CH₃)₃), 0.07 (s, 3H, SiCH₃), 0.17 (s, 3H, SiCH₃); ¹³C NMR (125 MHz;CDCl₃): δ 169.8, 169.2, 146.6, 137.3, 128.4, 128.0, 95.4, 73.8, 72.0,68.0, 67.8, 67.0, 66.9, 52.1, 32.1, 25.4, 20.9, 18.6, 17.8, 13.5, −4.7,−5.7; ESI-TOF HRMS m/z calcd for C₃₀H₅₂O₁₁PSi [M+H]⁺: 647.3017. found:647.3001.

Methyl2-O-acetyl-3-O-benzyl-4-O-tert-butyldimethylsilyl-α-L-idopyranosyluronate-(1→4)-tert-butyldimethylsilyl(2-azido-3-O-benzyl-6-O-levulinoyl-2-deoxy-β-D-glucopyranoside) (14)

Compound 6 (92 mg, 0.14 mmol) and 7 (77 mg, 0.17 mmol) wereco-evaporated with toluene (3.0×1.0 mL) and placed under vacuumovernight. The mixture was dissolved in CH₂Cl₂ (4.2 mL), and freshlyactivated 4 Å molecular sieves (0.21 g) were added. After stirring at rtfor 15 min, the temperature was lowered to −10° C. and the mixturestirred for an additional 15 min. TMSOTf (31 μL, 0.18 mmol) was addeddropwise to the reaction mixture. The reaction was stirred at −30° C.for 30 min, quenched with Et₃N (1.0 mL), filtered through a silica pad,and concentrated under reduced pressure. Silica gel flash chromatography(5:1→4:1 hexanes:EtOAc) afforded the desired product (120 mg) in 93%yield. ¹H NMR (600 MHz; CDCl₃): δ 7.37-7.24 (m, 10H, OCH₂Ph), 5.20 (d,J=4.2 Hz, 1H, H-1 of IdoA), 4.85 (t, J=4.1 Hz, 1H, H-2 of IdoA), 4.82(d, J=10.6 Hz, 1H, OCH₂Ph), 4.74 (d, J=11.8 Hz, 1H, OCH₂Ph), 4.71 (d,J=10.8 Hz, 1H, OCH₂Ph), 4.69 (d, J=3.9 Hz, 1H, H-5 of IdoA), 4.64 (d,J=11.8 Hz, 1H, OCH₂Ph), 4.54 (dd, J=11.8, 2.1 Hz, 1H, H-1 of GlcN),4.51-4.45 (m, 1H, H-6), 4.20-4.03 (m, 1H, H-6), 3.99 (t, J=4.3 Hz, 1H,H-4 of IdoA), 3.91-3.77 (m, 1H, H-4 of GlcN), 3.61 (t, J=4.3 Hz, 1H, H-3of IdoA), 3.54 (s, 3H, CO₂CH₃), 3.48 (ddd, J=9.8, 5.9, 2.2 Hz, 1H, H-5of GlcN), 3.37-3.24 (m, 2H, H-2 and H-3 of GlcN), 2.89-2.66 (m, 2H,COCH₂CH₂COCH₃), 2.61 (t, J=6.7 Hz, 2H, COCH₂CH₂COCH₃), 2.19 (s, 3H,COCH₂CH₂COCH₃), 2.00 (s, 3H, COCH₃), 0.92 (s, 9H, SiC(CH₃)₃), 0.81 (s,9H, SiC(CH₃)₃), 0.13 (d, J=4.6 Hz, 6H, SiCH₃), −0.06 (s, 3H, SiCH₃),−0.12 (s, 3H, SiCH₃); ¹³C NMR (125 MHz; CDCl₃): δ 206.5, 172.2, 170.2,169.9, 138.2, 137.7, 128.5, 128.1, 128.1, 127.9, 127.4, 97.6, 97.1,80.6, 76.8, 76.5, 75.4, 74.8, 73.4, 72.7, 71.5, 69.9, 68.9, 68.5, 62.6,51.7, 38.0, 29.9, 28.0, 25.6, 25.5, 20.9, 18.0, 17.8, −4.3, −4.7, −5.2,−5.5; ESI-TOF HRMS m/z calcd for C₄₆H₆₉N₃O₁₄Si₂ [M+Na]⁺: 966.4216.found: 966.4211.

Methyl2-O-acetyl-3-O-benzyl-4-O-tert-butyldimethylsilyl-α-L-idopyranosyluronate-(1→4)-2-azido-3-O-benzyl-1-O-(2-(2-((2S)-bicyclo[2.2.1]hept-5-en-2-yl-methoxy)ethoxy)ethyl)-6-O-levulinoyl-2-deoxy-α-D-glucopyranoside(5)

Compound 15 (37 mg, 60 μmol) and 8 (33 mg, 70 μmol) were co-evaporatedwith toluene (3.0×1.0 mL) and placed under vacuum overnight. The mixturewas dissolved in CH₂Cl₂ (1.7 mL) and freshly activated 4 Å molecularsieves (80 mg) were added. After stirring at rt for 15 min, thetemperature was lowered to −30° C. and the mixture stirred for anadditional 15 min. TMSOTf (14 μL, 70 μmol) was added to dropwise to thereaction mixture. The reaction was stirred at −10° C. for 10 min, slowlyraised to rt over 15 min, quenched with Et₃N (0.50 mL), filtered througha silica pad, and concentrated under reduced pressure. Silica gel flashchromatography (10:1→4:1→3:1 hexanes:EtOAc) afforded the desired product(35 mg) in 67% yield. ¹H NMR (500 MHz; CDCl₃): δ 7.38-7.25 (m, 10H,OCH₂Ph), 6.07 (ddd, J=25.6, 5.7, 3.0 Hz, 2H, CH═CH of Nb), 5.19 (d,J=4.2 Hz, 1H, H-1 of IdoA), 4.89-4.79 (m, 2H, H-2 of IdoA, OCH₂Ph),4.72-4.68 (m, 3H, H-5 of IdoA, OCH₂Ph), 4.64 (d, J=11.9 Hz, 1H, OCH₂Ph),4.52 (dd, J=12.1, 2.2 Hz, 1H, H-6 of GlcN), 4.34 (d, J=7.9 Hz, 1H, H-1of GlcN), 4.12 (dd, J=12.1, 2.2 Hz, 1H, H-6 of GlcN), 4.02-3.93 (m, 2H,H-4 of IdoA, OCH₂ of PEG linker), 3.87 (dd, J=9.8, 8.9 Hz, 1H, H-4 ofGlcN), 3.82-3.55 (m, 5H, H-3 of IdoA, OCH₂ of PEG linker), 3.54 (s, 3H,CO₂CH₃), 3.53-3.29 (m, 5H, H-5 of GlcN, H-2 of GlcN, H-3 of GlcN, OCH₂of PEG linker), 2.88-2.67 (m, 4H, CH—CH═CH of Nb, COCH₂CH₂COCH₃),2.67-2.56 (m, 2H, COCH₂CH₂COCH₃), 2.19 (s, 3H, COCH₂CH₂COCH₃), 2.00 (s,3H, OCOCH₃), 1.75-1.66 (m, 1H, CH of Nb), 1.39-1.15 (m, 4H, CH₂ of Nb),0.81 (s, 9H, SiC(CH₃)), −0.06 (s, 3H, SiCH₃), −0.11 (s, 3H, SiCH₃); ¹³CNMR (125 MHz; CDCl₃): δ 136.6, 128.5, 128.2, 127.8, 102.2, 80.9, 76.8,76.1, 75.0, 73.2, 72.9, 71.6, 70.7, 70.4, 66.0, 45.0, 43.6, 38.8, 38.0,29.8, 28.1, 25.5; ESI-TOF HRMS m/z calcd for C₅₂H₇₃N₃O₁₆Si [M+Na]⁺:1046.4658. found: 1046.4670

Methyl2-O-acetyl-3-O-benzyl-4-O-tert-butyldimethylsilyl-α-L-idopyranosyluronate-(1→4)-2-azido-3-O-benzyl-6-O-levulinoyl-2-deoxy-β-D-glucopyranosyltrichloroacetimidate (15)

Compound 14 (840 mg, 0.89 mmol) was dissolved in THF (27 mL) and thesolution was cooled to 0° C. 1M TBAF (1.2 mL, 1.2 mmol) and AcOH (60 μL,1.1 mmol) were added simultaneously, and the reaction was stirred for 30min at 0° C. The reaction was quenched with aqueous NaHCO₃ (10 mL),extracted with CH₂Cl₂ (2.0×10 mL), and subsequently washed with H₂O, 1MH₂SO₄, and then H₂O (10 mL for each wash). After concentrating underreduced pressure, the crude mixture (0.89 mmol) was dissolved in CH₂Cl₂(27 mL) and cooled to 0° C. To the reaction mixture,trichloroacetonitrile (1.3 mL, 13 mmol) and DBU (26 μL, 0.18 mmol) wereadded. The reaction was stirred at 0° C. for 12 h, quenched with Et₃N(1.0 mL), and concentrated under reduced pressure. Silica gel flashchromatography (5:1→4:1→3:1 hexanes:EtOAc+1% Et₃N) afforded the desiredproduct (770 mg) in 89% yield over two steps. ¹H NMR (600 MHz; CDCl₃): δ8.72 (s, 1H, OCNHCCl₃), 7.47-7.28 (m, 10H, OCH₂Ph), 6.37 (d, J=3.6 Hz,1H, H-1 of IdoA), 5.24 (d, J=4.7 Hz, 1H, H-1 of GlcN), 4.96 (d, J=10.5Hz, 1H, OCH₂Ph), 4.90 (t, J=4.4 Hz, 1H, H-2 of GlcN), 4.75 (d, J=11.7Hz, 1H, OCH₂Ph), 4.71 (d, J=10.5 Hz, 1H, OCH₂Ph), 4.66 (d, J=11.8 Hz,1H, OCH₂Ph), 4.63 (d, J=4.2 Hz, 1H, H-5 of GlcN), 4.52 (dd, J=12.3, 1.8Hz, 1H, H-6 of GlcN), 4.13 (dd, J=12.3, 4.3 Hz, 1H, H-6 of GlcN),4.08-3.98 (m, 3H, H-4 and H-5 of IdoA, H-4 of GlcN), 3.91 (dd, J=10.2,8.5 Hz, 1H, H-3 of IdoA), 3.69 (dd, J=10.2, 3.6 Hz, 1H, H-2 of IdoA),3.66-3.61 (m, 1H, H-3 of GlcN), 3.57 (s, 3H, CO₂CH₃), 2.89-2.69 (m, 2H,COCH₂CH₂COCH₃), 2.67-2.54 (m, 2H, COCH₂CH₂COCH₃), 2.18 (s, 3H,COCH₂CH₂COCH₃), 2.01 (s, 3H, COCH₃), 0.82 (d, J=2.5 Hz, 9H, SiC(CH₃)₃),−0.05 (s, 3H, SiCH₃), −0.09 (s, 3H, SiCH₃); ¹³C NMR (125 MHz; CDCl₃): δ206.5, 172.1, 170.2, 170.0, 160.7, 137.7, 137.6, 128.7, 128.2, 128.1,128.0, 127.9, 127.6, 97.7, 94.4, 78.2, 77.2, 76.7, 75.1, 75.0, 73.0,72.0, 71.9, 70.2, 69.0, 62.8, 61.9, 51.7, 38.0, 29.9, 28.0, 25.5, 20.1,17.8, 4.7, 5.4; ESI-TOF HRMS m/z calcd for C₄₂H₅₅N₃O₁₅SiCl₃ [M+Na]⁺:997.2366. found: 997.2415.

Example 2: Synthesis of Glycopolymers 1-4

An exemplary method of synthesizing glycopolymers (1-4) from compound 5is schematically illustrated in FIG. 3.

Methyl3-O-benzyl-4-O-tert-butyldimethylsilyl-α-L-idopyranosyluronate-(1→4)-2-amino-3-O-benzyl-1-O-(2-(2-((2S)-bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)ethyl)-2-deoxy-α-D-glu-copyranoside(16)

Compound 5 (17 mg, 20 μmol) was dissolved in anhydrous MeOH (0.80 mL),and 1,3-propanedithiol (0.14 mL, 60 μmol) and DIPEA (0.12 mL, 60 μmol)were added dropwise. Upon confirmation of partial disappearance of 5 byTLC, flame-dried K₂CO₃ (2.4 mg, 20 μmol) was added and the reactionmixture was stirred for 24 h at rt. The reaction was quenched with Dowex5W-X8 (H⁺ form), filtered through a pad of Celite, and concentratedunder reduced pressure. Silica gel flash chromatography (1:1hexanes:EtOAc) afforded the desired product (14 mg) in 93% yield. ¹H NMR(500 MHz; CDCl₃): δ 7.48-7.20 (m, 10H, OCH₂Ph), 6.17-5.88 (m, 2H, CH═CHof Nb), 5.25 (d, J=4.5 Hz, 1H, H-1 of IdoA), 4.96 (d, J=11.4 Hz, 1H,OCH₂Ph), 4.83 (d, J=11.4 Hz, 1H, OCH₂Ph), 4.68-4.51 (m, 3H, OCH₂Ph, H-5of IdoA), 4.32 (d, J=8.0 Hz, 1H, H-1 of GlcN), 4.09-3.76 (m, 5H, H-6 ofIdoA, H-4 of IdoA, H-2 of IdoA, OCH₂ of PEG linker), 3.76-3.53 (m, 10H,H-2 of IdoA, H-6 of IdoA, OCH₂ of PEG linker, CO₂CH₃), 3.47-3.32 (m, 4H,H-4 of GlcN, OCH₂ of PEG linker, H-5 of GlcN, H-3 of GlcN), 2.84-2.68(m, 3H, H-2 of GlcN, CH—CH═CH of Nb), 1.69-1.63 (m, 1H, CH of Nb),1.40-1.03 (m, 4H, CH₂ of Nb), 0.82 (s, 9H, SiC(CH₃)), −0.03 (d, J=7.3Hz, 7H, SiCH₃); ¹³C NMR (125 MHz; CD₃OD): δ 171.7, 140.1, 139.7, 137.7,137.5, 129.2, 128.8, 128.4, 104.5, 102.3, 84.1, 79.5, 77.7, 77.0, 75.2,74.4, 72.8, 71.5, 71.4, 61.8, 57.8, 52.4, 45.8, 44.9, 42.8, 40.1, 30.6,26.1, 18.7, −4.5, −5.1; ESI-TOF HRMS: m/z calcd for C₄₅H₆₆NO₁₃Si [M−H]⁻856.4303. found: 856.4326.

Methyl2-O-acetyl-3-O-benzyl-4-O-tert-butyldimethylsilyl-α-L-idopyranosyluronate-(1→4)-2-acetylamido-3-O-benzyl-1-O-(2-(2-((2S)-bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)eth-yl)-2-deoxy-α-D-glucopyranoside(17)

Compound 5 (190 mg, 0.18 mmol) was dissolved in anhydrous MeOH (11 mL).1,3-propanedithiol (1.1 mL, 5.4 mmol) and DIPEA (1.1 mL, 6.3 mmol) wereadded dropwise, and the reaction mixture was stirred for 24 h at rt. Thereaction was quenched with Dowex 5W-X8 (H⁺ form), filtered through a padof Celite, and concentrated under reduced pressure. Silica gel flashchromatography (30:2:1→20:2:1 EtOAc:MeOH:H₂O) afforded the desiredproduct (150 mg), and the resulting intermediate was dissolved inpyridine (2.8 mL). To this mixture, a solution of hydrazine monohydrate(1.2 mmol) and AcOH (9.9 mmol) in pyridine (17 mL) was added at rt. Thereaction mixture was diluted with CH₂Cl₂ (10 mL), washed with cold water(15 mL), saturated NaHCO₃ (15 mL), water (15 mL), and saturated brine(15 mL). The combined organic fractions were dried over MgSO₄ andconcentrated under reduced pressure. Silica gel flash chromatography(20:2:1 EtOAc:MeOH:H₂O) afforded the desired product (140 mg) in 87%yield over two steps. ¹H NMR (500 MHz; CDCl₃): δ 7.46-7.27 (m, 10H,OCH₂Ph), 6.14-5.98 (m, 2H, CH═CH of Nb), 5.31 (d, J=4.4 Hz, 1H, H-1 ofIdoA), 4.99 (d, J=11.2 Hz, 1H, OCH₂Ph), 4.88 (q, J=3.6 Hz, 1H, H-2 ofIdoA), 4.80-4.68 (m, 2H, OCH₂Ph, H-5 of IdoA), 4.67-4.55 (m, 2H,OCH₂Ph), 4.36 (dt, J=8.0, 4.1 Hz, 1H, H-1 of GlcN), 4.05-3.85 (m, 4H,H-4 of IdoA, H-6 of GlcN, H-5 of GlcN, OCH₂ of PEG linker), 3.85-3.34(m, 15H, OCH₂ of PEG linker, H-6 of GlcN, H-3 of IdoA, CO₂CH₃, H-3 ofGlcN, H-4 of GlcN), 2.89 (dd, J=10.0, 7.8 Hz, 1H, H-2 of GlcN),2.81-2.67 (m, 2H, CH—CH═CH of Nb), 2.01 (s, 3H, OCOCH₃), 1.42-1.14 (m,5H, CH and CH₂ of Nb), 0.82 (s, 9H, SiC(CH₃)), −0.08 (d, J=7.3 Hz, 6H,SiCH₃); ¹³C NMR (125 MHz; CDCl₃): δ 170.2, 138.7, 137.8, 136.7, 128.5,128.1, 127.9, 127.6, 97.8, 76.8, 76.2, 75.7, 75.5, 74.2, 72.7, 72.0,70.7, 70.4, 69.3, 69.0, 61.7, 56.6, 51.9, 45.1, 43.8, 41.7, 38.9, 38.6,29.9, 25.7, 21.1, 17.9, −4.5, −5.4; ESI-TOF HRMS: m/z calcd forC₄₇H₇₀NO₁₄Si [M−H]⁻ 900.4565. found: 900.4568.

Methyl3-O-benzyl-2-O-sulfonato-4-O-tert-butyldimethylsilyl-α-L-idopyranosyluronate-(1→4)-3-O-benzyl-1-O-(2-(2-((2S)-bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)ethyl)-2-deoxy-2-sulfonatamido-6-O-sulfonato-α-D-glucopyranoside(18)

Compound 16 (9.2 mg, 10 μmol) was dissolved in freshly distilledpyridine (1.0 mL) and to this SO₃.Py (50 mg, 0.32 mmol) and Et₃N (0.20mL) were added. The reaction mixture was stirred at rt for 24 h,refluxed at 50° C. for 24 h, quenched with MeOH (1.0 mL), andconcentrated to afford a golden syrup. Purification by Sephadex LH-20gel filtration (1:1 CH₂Cl₂:MeOH), followed by silica gel flashchromatography (15:2:1→10:2:1→8:2:1 EtOAc:MeOH:H₂O) gave the desiredproduct (8.7 mg) in 78% yield. ¹H NMR (500 MHz; CD₃OD): δ 7.51-7.50 (m,2H, OCH₂Ph), 7.43-7.41 (m, 2H, OCH₂Ph), 7.37-7.34 (m, 2H, OCH₂Ph),7.30-7.26 (m, 3H, OCH₂Ph), 7.23-7.22 (m, 1H, OCH₂Ph), 6.11-6.04 (m, 2H,CH═CH of Nb), 5.30 (s, H-1 of IdoA), 4.98 (d, J=12.5 Hz, 1H, OCH₂Ph),4.87 (d, J=11.5 Hz, 1H, OCH₂Ph), 4.78 (d, J=6 Hz, 1H, H-1 of GlcN), 4.67(d, J=11.5 Hz, 1H, OCH₂Ph), 4.59 (d, J=12.5 Hz, 1H, OCH₂Ph), 4.43 (s,1H, H-2 of IdoA), 4.37-4.28 (m, 2H, H-6, H-6 of GlcN), 4.13-4.12 (m, 1H,H-4 of GlcN), 4.05-4.03 (m, 1H, H-5 of GlcN), 3.96-3.93 (m, 1H, H-4 ofIdoA), 3.81-3.78 (m, 2H, H-3 of IdoA, OCH₂ of PEG linker), 3.73-3.71 (m,2H, OCH₂ of PEG linker), 3.69-3.56 (m, 5H, OCH₂ of PEG linker),3.54-3.51 (m, 1H, H-2 of GlcN), 3.45-3.35 (m, 2H, OCH₂ of PEG linker),3.15 (s, 3H, CO₂CH₃), 2.77 (s, 1H, CH—CH═CH of Nb), 2.72 (s, 1H,CH—CH═CH of Nb), 2.11 (s, 3H, OCOCH₃), 1.98 (s, 3H, OCOCH₃), 1.72-1.68(m, 1H, CH of Nb), 1.38-1.21 (m, 3H, CH₂ of Nb), 1.15-1.12 (m, 1H, CH₂of Nb), 0.76 (s, 9H, SiC(CH₃)), −0.17 (s, 3H, SiCH₃), −0.24 (s, 3H,SiCH₃); ¹³C NMR (125 MHz; CD₃OD): δ 172.8, 141.1, 140.4, 138.7, 131.6,131.0, 130.6, 130.4, 130.3, 130.2, 130.0, 129.4, 104.3, 100.1, 80.9,78.0, 77.0, 72.7, 72.4, 71.0, 70.7, 56.2, 53.5, 46.9, 46.0, 43.8, 41.1,31.7, 27.4, 19.9, −3.0, −4.4; ESI-TOF HRMS: m/z calcd for C₄₅H₆₆NO₂₂S₃Si[M−H]⁻ 1016.2597. found: 1016.2583.

Methyl2-O-sulfonato-3-O-benzyl-4-O-tert-butyldimethylsilyl-α-L-idopyranosyluronate-(1→4)-2-acetylamido-3-O-benzyl-1-O-(2-(2-((2S)-bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)ethyl)-2-deoxy-6-O-sulfonato-α-D-glucopyranoside(19)

To a solution of compound 16 (130 mg, 0.15 mmol) in anhydrous MeOH (8.4mL) at ambient temperature were added Ac₂O (0.30 mL, 3.0 mmol) and Et₃N(0.50 mL). Additional amounts of Ac₂O (0.30 mL, 3.0 mmol) were addedevery hour until complete conversion to the desire product was observedby TLC (at least 4 h). The reaction mixture was directly loaded onto aSephadex LH-20 gel filtration column and eluted with 1:1 CH₂Cl₂:MeOH.The N-acetylated intermediate was dissolved in freshly distilledpyridine (8.1 mL), and SO₃—Py (450 mg, 3.3 mmol) and Et₃N (1.6 mL) wereadded. The reaction mixture was stirred at rt for 24 h, refluxed at 50°C. for 24 h, quenched with MeOH (5.0 mL), and concentrated to afford agolden syrup. Purification by Sephadex LH-20 gel filtration (1:1CH₂Cl₂:MeOH), followed by silica gel flash chromatography (10:2:1EtOAc:MeOH:H₂O) gave the desired product (130 mg) in 85% yield over twosteps. ¹H NMR (500 MHz; CD₃OD): δ 7.53-7.10 (m, 10H, OCH₂Ph), 6.12-5.95(m, 2H, CH═CH of Nb), 5.35 (s, 1H, H-1 of IdoA), 4.84 (m, 2H, H-5 ofIdoA, OCH₂Ph), 4.73 (d, J=11.2 Hz, 1H, OCH₂Ph), 4.60 (d, J=11.6 Hz, 1H,OCH₂Ph), 4.55 (d, J=8.3 Hz, 1H, H-1 of GlcN), 4.52 (dd, J=2.1, 1.1 Hz,1H, H-2 of IdoA), 4.47 (d, J=11.2 Hz, 1H, OCH₂Ph), 4.41 (dd, J=11.3, 2.2Hz, 1H, OCH₂ of PEG linker), 4.29 (dd, J=11.2, 5.0 Hz, 1H, OCH₂ of PEGlinker), 4.00-3.87 (m, 4H, H-4 of IdoA, H-2 of GlcN, H-3 of GlcN, H-6 ofGlcN), 3.86 (t, J=1.8 Hz, 1H, H-3 of IdoA), 3.79-3.48 (m, 11H, H-6 ofGlcN, OCH₂ of PEG linker, H-4 of GlcN, H-5 of GlcN), 3.46-3.36 (m, 1H,OCH₂ of PEG linker), 3.33 (s, 3H, CO₂CH₃), 2.74 (d, J=31.6 Hz, 2H,CH—CH═CH of Nb), 1.85 (d, J=1.2 Hz, 3H, NHCOCH₃), 1.67 (d, J=4.5 Hz, 1H,CH of Nb), 1.39-1.04 (m, 4H, CH₂ of Nb), 0.78 (d, J=1.2 Hz, 9H,SiC(CH₃)), −0.11 (dd, J=47.6, 1.1 Hz, 6H, SiCH₃); ¹³C NMR (125 MHz;CD₃OD): δ 173.3, 172.1, 139.7, 139.2, 137.7, 137.5, 129.9, 129.5, 129.1,128.6, 102.6, 99.2, 82.3, 77.1, 76.2, 75.7, 75.3, 74.1, 72.8, 71.5,71.4, 70.1, 69.7, 67.8, 56.5, 52.4, 45.9, 44.9, 42.8, 40.0, 30.7, 26.2,23.0, 18.9, −4.2, −5.4; ESI-TOF HRMS: m/z calcd for C₄₇H₆₇NO₂₀NaSiS₂[M+Na]⁺ 1080.3365. found: 1080.3392.

Methyl3-O-benzyl-4-O-tert-butyldimethylsilyl-α-L-idopyranosyluronate-(1→4)-2-acetyl-amido-3-O-benzyl-1-O-(2-(2-((2S)-bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)ethyl)-2-de-oxy-α-D-glucopyranoside(20)

To a solution of compound 16 (130 mg, 0.15 mmol) in anhydrous MeOH (8.4mL) at ambient temperature were added Ac₂O (0.30 mL, 3.0 mmol) and Et₃N(0.50 mL). Additional amounts of Ac₂O (0.30 mL, 3.0 mmol) were addedevery hour until complete conversion to the desired product was observedby TLC (at least 4 h). Purification by Sephadex LH-20 gel filtration(1:1 CH₂Cl₂:MeOH), followed by silica gel flash chromatography (20:2:1EtOAc:MeOH:H₂O) gave the desired product (140 mg) in quantitative yield.¹H NMR (600 MHz; CDCl₃): δ 7.36-7.09 (m, 10H, OCH₂Ph), 6.12-5.87 (m, 2H,CH═CH of Nb), 5.11 (s, 1H, H-1 of IdoA), 4.76-4.72 (m, 1H, H-1 of GlcN),4.70 (t, J=2.7 Hz, 1H, H-4 of IdoA), 4.68-4.56 (m, 2H, OCH₂Ph),4.51-4.43 (m, 2H, OCH₂Ph), 4.00-3.92 (m, 1H, H-3 of IdoA), 3.91-3.80 (m,2H, OCH₂ of PEG linker, H-6 of GlcN), 3.80-3.17 (m, 19H, H-6 of GlcN,H-2 of GlcN, H-3 of GlcN, H-4 of GlcN, H-5 of GlcN, H-2 of IdoA, H-5 ofIdoA, OCH₂ of PEG linker, CO₂CH₃), 2.75-2.46 (m, 2H, CH—CH═CH of Nb),1.67 (d, J=4.2 Hz, 3H, NHCOCH₃), 1.64-1.53 (m, 1H, CH of Nb), 1.30-1.13(m, 4H, CH₂ of Nb), 0.71 (s, 9H, SiC(CH₃)), −0.15 (d, J=9.3 Hz, 6H,SiCH₃); ¹³C NMR (125 MHz; CDCl₃): δ 170.4, 169.7, 138.6, 137.5, 136.8,136.6, 128.8, 128.4, 128.2, 127.4, 107.3, 101.6, 100.9, 78.7, 77.0,76.4, 75.7, 75.5, 72.6, 72.5, 71.0, 70.7, 70.4, 69.8, 69.1, 68.8, 67.2,62.7, 52.1, 45.2, 43.8, 41.7, 38.8, 30.0, 29.9, 25.6, 23.3, 17.9, −4.7,−5.4; ESI-TOF HRMS: m/z calcd for C₄₇H₆₉NaNO₁₄Si [M+Na]⁺ 922.4380.found: 922.4385.

Methyl2-O-acetyl-3-O-benzyl-4-O-tert-butyldimethylsilyl-α-L-idopyranosyluronate-(1→4)-3-O-benzyl-1-O-(2-(2-((2S)-bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)ethyl)-2-deoxy-2-sulfonatamido-6-O-sulfonato-α-D-glucopyranoside(21)

To a solution of compound 17 (22 mg, 0.020 mmol) in freshly distilledpyridine (2.3 mL) were added SO₃.Py (110 mg, 0.60 mmol) and Et₃N (0.5mL). The reaction mixture was stirred at rt for 24 h and refluxed at 50°C. for 24 h, quenched with MeOH (1.0 mL), and concentrated to afford agolden syrup. Purification by Sephadex LH-20 gel filtration (1:1CH₂Cl₂:MeOH), followed by silica gel flash chromatography (15:2:1→10:2:1EtOAc:MeOH:H₂O) gave the desired product (26 mg) in 78% yield. ¹H NMR(600 MHz; CD₃OD): δ 7.79-7.03 (m, 10H, OCH₂Ph), 6.25-5.94 (m, 2H, CH═CHof Nb), 5.31 (d, J=4.4 Hz, 1H, H-1 of IdoA), 5.05-4.76 (m, 4H, H-2 ofIdoA, H-5 of IdoA, OCH₂Ph), 4.75-4.64 (m, 1H, OCH₂Ph), 4.65-4.47 (m, 2H,OCH₂Ph, H-1 of GlcN), 4.38 (m, 1H, H-6 of GlcN), 4.20 (m, 1H, H-6 ofGlcN), 4.07-3.85 (m, 4H, H-2 of IdoA, H-2 of GlcN, H-2 of GlcN, OCH₂ ofPEG linker), 3.86-3.71 (m, 1H, OCH₂ of PEG linker), 3.71-3.45 (m, H,11H, H-3 of IdoA, H-3 of GlcN, H-5 of GlcN, OCH₂ of PEG linker), 3.40(s, 3H, CO₂CH₃), 2.81-2.74 (m, 2H, CH—CH═CH of Nb), 2.07 (s, 3H,OCOCH₃), 1.52-1.10 (m, 5H, CH and CH₂ of Nb), 0.83 (s, 9H, SiC(CH₃)),−0.09 (s, 3H, SiCH₃), −0.13 (s, 3H, SiCH₃); ¹³C NMR (125 MHz; CDCl₃): δ173.3, 172.0, 171.8, 139.8, 139.1, 137.7, 137.5, 102.8, 99.2, 82.1,79.8, 77.1, 76.6, 76.2, 75.9, 75.3, 75.2, 73.2, 71.5, 71.4, 71.3, 71.0,70.1, 69.8, 69.4, 67.3, 56.3, 52.4, 45.9, 44.9, 42.8, 40.0, 37.4, 36.0,30.7, 23.0, 21.2, 18.7, −4.3, −5.4; ESI-TOF HRMS: m/z calcd forC₄₇H₆₇NO₂₀NaSiS₂ [M+Na]⁺ 1080.3365. found: 1080.3392.

Protected HS Glycopolymers (22-25)

Monomers 18-21 were converted into polymers 22-25, which contain thefollowing functional groups: R₁═SO₃ ⁻, R₂═SO₃ ⁻, R₃═SO₃ ⁻ (1); R₁═SO₃ ⁻,R₂═SO₃ ⁻, R₃═Ac⁻ (2); R₁═H, R₂═H, R₃═Ac (3); R₁═H, R₂═SO₃ ⁻, R₃═SO₃ ⁻(4). In a typical polymerization, a small vial was charged with monomer(18-21; 6.0 mg, 5.0 μmol) and a small stir bar under the flow of argon.To this was added degassed dichloroethane (DCE)/MeOH (10:1, 0.025 M) andbis-pyridine Grubbs catalyst ((H₂IMes)(Py)₂(Cl)₂Ru═CHPh)⁴ in DCE (5mg/mL stock solution, 24 μL, 0.11 μmol) by syringe at rt. The reactionmixture was stirred at rt for 1 h, quenched with ethyl vinyl ether (0.10mL), and diluted with diethyl ether (1.0 mL) and hexanes (0.50 mL) toobtain a white precipitate. The mixture was centrifuged to remove theorganic layer, and the resulting white solid (83-98% conversion) wasdried in vacuo. ¹H NMR confirmed disappearance of the norborneneolefinic peaks at 6.04-6.11 ppm. The protected polymers werecharacterized by size exclusion chromatography multi-angle lightscattering (SEC-MALS) using a system equipped with an MZ-Gel SDplusorganic column (10E5 Å, MZ Analysentechnik), a light scattering detector(miniDAWN, Wyatt Technology), and a refractive index detector (TREOS,Wyatt Technology), and 0.2 M LiBr in DMF as the mobile phase. ¹H NMR(500 MHz; D₂O): δ 7.49-7.15 (m, 10H), 5.42 (br, 1H), 5.18 (br, 1H), 4.75(br, 1H), 4.65-4.51 (m, 2H), 4.38 (br, 1H), 4.05-3.84 (m, 4H), 3.86 (br,1H), 3.79-3.50 (m, 13H), 3.46 (br, 1H), 3.31 (br, 3H) 3.30-3.25 (m, 2H),2.49 (br, 2H), 1.94 (br, 3H), 1.79-1.07 (m, 5H), 0.77 (br, 9H), −0.07(br, 3H), −0.17 (br, 3H).

2-O-sulfonato-α-L-idopyranosyluronate-(1→4)-1-O-(2-(2-((2S)bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)ethyl)-2-deoxy-2-sulfonatamido-6-O-sulfonato-α-D-glucopyranoside(26)

Compound 18 (14 mg, 0.013 mmol) was dissolved in THF (1.3 mL), and TMSOK(33 mg, 0.26 mmol) was added to the reaction mixture. The reaction wasstirred for 24 h at rt and quenched with MeOH (1.0 mL). The crudereaction mixture was loaded directly onto a Sephadex G-25 gel filtrationcolumn and eluted with 100% H₂O, and fractions were combined,lyophilized, and subjected to hydrogenation. The intermediate wasdissolved in a 3:2 mixture of 80 mM phosphate buffered saline (2.4 mL,pH=7.0) and MeOH (0.80 mL). To this, Pd(OH)₂/charcoal (80 mg, 8× byweight of starting material) was added, and the reaction was carried outunder 1 atm H₂ gas for 3 d. The reaction mixture was filtered using avacuum filtration system (0.45 μm PES membrane, VWR) and desalted on aSephadex G-25 column in 100% H₂O to obtain the desired product 60% yieldafter lyophilization. ¹H NMR (500 MHz, D₂O) δ 5.24 (s, 1H, H-1 of IdoA),4.83 (d, J=2.7 Hz, 1H, H-5 of IdoA), 4.73 (d, J=8.2 Hz, 1H, H-1 ofGlcN), 4.44 (m, 1H, H-6 of GlcN), 4.34 (m, 2H, H-6 of GlcN, H-2 ofIdoA), 4.10 (m, 2H, H-3 of IdoA, H-4 of IdoA), 4.01-3.71 (m, 11H, OCH₂of PEG linker, H-3 of GlcN, H-4 of GlcN, H-5 of GlcN), 3.44-3.31 (m, 2H,OCH₂ of PEG linker), 3.16 (m, 1H, H-2 of GlcN), 2.28-2.20 (m, 2H,bridgehead CH₂ of Nb), 1.82 (s, 2H, CH₂ of Nb), 1.59 (s, 2H, CH₂ of Nb),1.48-1.36 (m, 2H, CH of Nb), 1.27-1.19 (m, 2H, CH₂ of Nb), 1.08 (s, 1H,CH of Nb).

Deprotected HS Glycopolymers (1-4)

Polymers 22-25 were deprotected to obtain final polymers 1-4, whichcontain the following functional groups: R₁═SO₃ ⁻, R₂═SO₃ ⁻, R₃═SO₃ ⁻(1); R₁═SO₃ ⁻, R₂═SO₃ ⁻, R₃═Ac⁻ (2); R₁═H, R₂═H, R₃═Ac (3); R₁═H, R₂═SO₃⁻, R₃═SO₃ ⁻ (4). In a typical reaction, polymer (11 mg, 10 μmol perunit) was dissolved in THF (1.0 mL), and TBAI (7.0 mg, 20 μmol) andTMSOK (25 mg, 0.20 mmol) were added. The reaction was stirred for 24 hat rt and quenched with MeOH (1.0 mL). The crude reaction mixture wasloaded directly onto a Sephadex G-25 gel filtration column and elutedwith 100% H₂O. The polymer-containing fractions were combined,lyophilized, and subjected to hydrogenation. In a typical hydrogenationreaction, the polymer from the previous reaction was dissolved in a 3:2mixture of 80 mM phosphate buffered saline (0.9 mL, pH=7.0) and MeOH(0.60 mL). To this, Pd(OH)₂/charcoal (84 mg, 8× weight of polymer) wasadded, and the reaction was carried out under 1 atm H₂ gas for 3 d.Samples were filtered using a vacuum filtration system (0.45 μm PESmembrane, VWR) and desalted on a Sephadex G-25 column in 100% H₂O toobtain the desired polymers in 35-55% yield after lyophilization. ¹H NMRshowed disappearance of the benzyl and methyl ester peaks at 7.79-7.03ppm and 3.40 ppm, respectively. Deprotected polymers were characterizedby SEC-MALS using a system equipped with an OHpak water column (SB-804HQ, Shodex), a light scattering detector (miniDAWN, Wyatt Technology),and a refractive index detector (TREOS, Wyatt Technology), and 3 mM NaN₃and 6 mM NaNO₃ in H₂O as the mobile phase. ¹H NMR (500 MHz; D₂O): δ 5.03(br, 1H), 4.44 (br, 1H), 4.25-4.20 (m, 1H), 4.18-4.11 (m, 2H), 3.92 (br,1H), 3.84 (br, 2H), 3.75-3.40 (m, 12H), 3.34 (br, 1H), 3.19 (br, 1H),1.91 (br, 3H), 1.72 (br, 1H), 1.48-0.88 (m, 6H).

Properties of synthesized glycopolymers (1-4) are listed in Table 2:wherein the number average molecular weight (M_(n)) and polydispersityindex (PDI) were determined by size exclusion chromatography multi-anglelight scattering (SEC-MALS).

TABLE 2 Properties of Glycopolymers mol % M_(n) n Polymer MonomerCatalyst (g/mol) PDI (DP) 1 22 5 27,870 1.22 35 2 23 5 36,490 1.02 48 324 5 53,580 1.16 90 4 25 5 32,880 1.03 46

Example 3: Direct and Competitive Enzyme-Linked Immunosorbent Assay(ELISA)

A 96-well heparin-binding plate (BD Biosciences) was coated with 25μg/mL of heparin (Neoparin) for 12 h at room temperature. Wells wererinsed with phosphate-buffered saline (PBS) and blocked with 10% fetalbovine serum (FBS) in PBS for 1 h at 37° C. For the direct ELISA,various concentrations of RANTES (0.50-1024 nM; R&D Systems) wereserially diluted in 1% BSA in PBS and incubated in each well for 1.5 hat 37° C. For the competitive ELISA, RANTES (at 12 nM, thepre-determined EC50) was pre-incubated (3 h, 37° C.) with variousconcentrations of heparin (0.010-40 μg/mL) or glycopolymers 1-4(0.10-180 gg/mL), and the co-mixture was added to the 96-well plate for1.5 h at 37° C. Wells were washed three times with PBST (PBS+0.1%Tween-20), incubated with a mouse anti-RANTES antibody (R&D Systems) for1 h at 37° C., washed three times with PBST, and incubated with ahorseradish peroxidase (HRP)-conjugated anti-mouse IgG antibody (GEHealthcare Life Sciences) for 1 h at 37° C. After three washes withPBST, RANTES binding was detected using a 3,3′,5,5′-tetramethylbenzidine(TMB) substrate kit (Thermo Scientific) according to the manufacturer'sinstructions. Fluorescence was measured at 450 nm using a Victor 3 platereader (PerkinElmer). The half-maximal effective concentration (EC50)and half maximal inhibitory concentration (IC50) were calculated usingKaleidaGraph software (Synergy). IC50 values reported in the paper arefor both the mass and molar concentrations of antagonist. IC50 valueswere also corrected for ligand valence (Table 3) by calculating the masspercentage of the disaccharide epitope contributing to eachdisaccharide-norbornyl linker unit, and then dividing by the molecularweight of the disaccharide epitope.

TABLE 3 Half Maximal Inhibitory Concentration (IC50) for Antagonists ofRANTES IC50 IC50 IC50 IC50 (μg/mL (μM Antagonist (μg/mL) (nM) of disac)of disac) 1 9.3 ± 1.1 334 ± 39  6.8 ± 0.80 11.7 ± 1.4 2 31.1 ± 6.2   852± 170 21.8 ± 4.4 40.3 ± 8.0 4 58.0 ± 5.7  1760 ± 170 40.6 ± 4.0 81.3 ±8.0 Heparin 0.90 ± 0.03 45.0 ± 1.5  0.90 ± 0.03  1.50 ± 0.05

Example 4: Chromogenic Assay for the Measurement of AntithrombinActivity

Factor Xa Activity:

The BIOPHEN Heparin Anti-Xa (2 stages) USP/EP kit (Aniara) was used todetermine factor Xa activity. This chromogenic anti-Xa method formeasuring homogeneous heparin in purified systems is in compliance withPharmacopoeias (USP, EP) and FDA guidelines. All reagents were preparedaccording to manufacturer's instructions and incubated at 37° C. for 15min. Varying concentrations of heparin (Neoparin) or glycopolymers 1-4(40 μL) and antithrombin (40 μL) were added to a microcentrifuge tube,mixed, and incubated at 37° C. for 2 min. Factor Xa (40 μL) was added tothe solution, incubated at 37° C. for exactly 2 min, and then the factorXa chromogenic substrate (40 μL) was added. After 2 min, the reactionwas quenched with citric acid (20 g/L, 240 μL), and the absorbance wasmeasured at 405 nm. The sample blank was obtained by mixing the reagentsin reverse order, and the resulting value was deducted from theabsorbance values measured in the assay.

Thrombin (Factor IIa) Activity:

The BIOPHEN Heparin Anti-IIa (2 stages) USP/EP kit (Aniara) was used todetermine factor IIa activity. This chromogenic anti-IIa method wasconducted according to the same procedure used for factor Xa.

The relative ability of each glycopolymer to block RANTES binding toheparin (EC50=12.2 nM) was tested by a competitive enzyme-linkedimmunosorbent assay. As shown in FIG. 4 and Table 3, trisulfatedglycopolymer 1 bound strongly to RANTES (IC50=9.3±1.1 μg/mL (334±39nM)), albeit with reduced affinity compared to heparin of similar chainlength (IC50=0.90±0.03 μg/mL (45.0±1.5 nM). However, the glycopolymercompeted more effectively for RANTES binding compared to heparin at itsmaximum inhibitory concentration. Whereas heparin exhibited a maximuminhibition of 58.4%, glycopolymer 1 inhibited RANTES binding by up to90.8% under the same assay conditions.

The effects of site-defined modifications to the sulfation pattern ofglycopolymer 1 on its affinity for RANTES was tested. Removal of eitherthe N-sulfate group of GlcN (glycopolymer 2) or the 2-O-sulfate group ofIdoA (glycopolymer 4) decreased binding to RANTES (IC50=31.1±6.2 μg/mL(852±170 nM) and 58.0±5.7 μg/mL (1760±170 nM), respectively), andunsulfated glycopolymer 3 had no appreciable activity (FIG. 4a ). Theobservation suggests that both of the sulfation pattern and sulfationdegree are important for determining the affinity of the glycopolymersfor RANTES. Importantly, none of the glycopolymers possessedanti-coagulant activity, as demonstrated by their inability topotentiate the inhibition of factor Xa and thrombin substrates byantithrombin III (FIG. 4b ). Thus, controlling the positioning ofsulfate groups within the glycopolymer enables the anti-inflammatoryfunction of HS/heparin to be dissected from its anti-coagulant function.Furthermore, modifications to the sulfation pattern can be exploited toadjust the affinity of the glycopolymers for different HS-bindingproteins and may facilitate the development of glycosaminoglycan-basedtherapeutic agents with fewer off-target side effects.

Example 5: Cell Migration Assay

L1.2 cells (mouse pre-B lymphocytes) stably transfected with CCR3, CCR5,or vector only, were kindly provided by Dr. Osamu Yoshie (KinkiUniversity, School of Medicine, Japan). Cells were maintained in RPMI1640 (Invitrogen) supplemented with 10% FBS, 100 μg/mLpenicillin/streptomycin (Invitrogen), and 50 μM 2-mercaptoethanol (SigmaAldrich). Cells were routinely analyzed by flow cytometry (FACSCalibur,Beckman Dickenson) to verify that cultures expressed adequate levels ofchemokine receptor (>90%) for migration and cell binding assays.

Experiments were performed using ChemoTx chambers (Neuroprobe). L1.2cells (wild-type or stably-transfected with CCR3 or CCR5) were harvestedand washed twice in flow cytometry buffer (Hank's Balanced Salt Solution(HBSS) with 2.5 mg/mL bovine serum albumin (BSA) and 10 mM HEPES). HumanRANTES (R&D Systems) was serially diluted in flow cytometry buffer(0.5-1024 nM), and 30 μL of each dilution was added to the bottom wellsof the ChemoTx chamber. Alternatively, in competitive migration assays,1 or 10 nM of RANTES was pre-incubated with various concentrations ofheparin or glycopolymer 1 (0.020-4.0 μg/mL) for 30 min at rt, and thesame volume of each solution was added to the bottom wells. The sampleplate was fitted with a 5-μm pore filter, and 106 cells (50 μL) wereplaced on top of each well. Cells were allowed to migrate through thefilter for 4 h at 37° C. and 5% CO2. Subsequently, non-migrating cellswere removed from the top of the filter by manual scraping; cellsadhering to the filter were dislodged using 20 μL of 2.5 mM EDTA for 30min at rt. Migrated cells were transferred (500×g, 5 min) to a 96-wellblack-walled clear-bottomed plate (Corning) using a funnel plate(Neuroprobe). Cells were lysed at −80° C. and stained with CyQUANT dye(Invitrogen) as described in the product literature. Fluorescence wasmeasured at 535 nm using a Victor 3 plate reader (PerkinElmer).

Example 6: Chemokine Cell Binding Assay

3×10⁶ L1.2 cells (wild-type or stably-transfected with CCR3) were washedtwice with flow cytometry buffer and incubated with RANTES (100 nM inflow cytometry buffer) for 45 min at rt. Alternatively, cells wereincubated with RANTES (100 nM in flow cytometry buffer) previouslytreated with various concentrations of heparin or glycopolymer 1 (0.02-2μg/mL) for 30 min at rt. Cells were spun twice (500×g, 5 min) through100% FBS (1.0 mL) to remove excess reagent and stained withphycoerythrin (PE)-conjugated anti-RANTES (1 test) in FACS buffer (100μL) for 1 h at 4° C. Cells were again spun twice through 100% FBS (1.0mL) and resuspended in flow cytometry buffer (500 μL) for flow cytometryanalysis. Immediately before analysis, 7-amino-actinomycin-D (7-AAD, 5μL, eBioscience) was added to evaluate cell viability. Cells wereanalyzed for PE intensity on a FACSCalibur flow cytometer (BeckmanDickenson, Caltech Flow Cytometry Facility) with 10,000 cell events persample. Data analysis was performed using FlowJo (Tree Star Inc.).

The chemotactic activity of RANTES, which is essential to thepathogenesis of allergic inflammatory responses such as asthma, ismediated in part by the G-protein coupled receptor CCR3. To test whetherglycopolymer 1 can interfere with RANTES-induced chemotaxis via CCR3, weprobed the migration of murine L1.2 pre-B cells that were stablytransfected with the CCR3 receptor. Using a modified Boyden chamber, weobserved that the directional migration of L1.2-CCR3 cells (but notwild-type L1.2 cells lacking CCR3) was dependent on the RANTESconcentration and elicited a maximal response at 10 nM (FIG. 5).Pre-incubation of RANTES (10 nM) with either glycopolymer 1 or heparindiminished the chemotactic activity of RANTES in a dose-dependent manner(FIG. 6). Further corroborating these results, lower levels of RANTESwere detected on the surface of CCR3-expressing cells after thechemokine (100 nM) was pretreated with glycopolymer 1 or heparin, asdetermined by flow cytometry analysis (FIG. 7).

Example 7: Synthesis and Characterization of Compound 31 andIntermediates Thereof

Compounds to be used in the further preparing procedures may besynthesized by a method shown in FIG. 10.

tert-Butyldimethylsilyl6-O-acetyl-2-azido-3-O-benzyl-4-tert-butyldimethylsilyl-2-deoxy-β-D-glucopyranoside(S2)

Compound S2 was prepared from6-O-acetyl-2-azido-3-O-benzyl-4-tert-butyldimethylsilyl-2-deoxy-β-D-glucopyranoside(S1) using a number of procedures. (See e.g., Orgueira H. A. et al.,Chem. Eur. J. 2003, 9, 140-169.) The analytical data were in agreementwith the reported spectra.

tert-Butyldimethylsilyl6-O-acetyl-2-azido-3-O-benzoyl-4-tert-butyldimethylsilyl-2-deoxy-β-D-glucopyranoside(S3)

Oxidation conditions were adapted from previously published procedures.Compound S2 (200 mg, 0.353 mmol) was added to a mixture of 1.00 mL ofcarbon tetrachloride, 1.00 mL of acetonitrile, and 1.50 mL of H₂O. Tothis, sodium metaperiodate (755 mg, 10 eq) and ruthenium dioxide (47.0mg, 1 eq) were added sequentially, and the reaction was stirred for 18 hat rt in the dark. The resulting slurry as diluted with CH₂Cl₂, and theaqueous layer was extracted with CH₂Cl₂ (3×). The organic layers werecombined, filtered through a pad of Celite, and concentrated in vacuo.The residue was purified by silica gel flash chromatography (20:1hexanes:EtOAc) to deliver 164 mg (80%) of S3 as a colorless oil. ¹H NMR(300 MHz; CDCl₃): δ 8.07 (d, J=7.2 Hz, 2H), 7.59 (t, J=7.5 Hz, 1H), 7.47(t, J=7.5 Hz, 2H), 5.16 (dd, J=10.2, 9.0 Hz, 1H), 4.72 (d, J=7.5 Hz,1H), 4.43 (dd, J=11.7, 1.8 Hz, 1H), 4.12 (dd, J=12.0, 6.3 Hz, 1H), 3.85(t, J=9.0 Hz, 1H), 3.61-3.56 (m, 1H), 3.44 (dd, J=10.5, 7.8 Hz, 1H),2.10 (s, 3H), 0.93 (s, 9H), 0.75 (s, 9H), 0.16 (s, 6H), 0.02 (s, 3H),−0.20 (s, 3H); ¹³C NMR (75 MHz; CDCl₃: δ 170.6, 165.4, 133.3, 129.9,128.5, 97.1, 74.9, 74.4, 69.6, 66.8, 63.0, 25.5, 22.2, 20.8, 17.9, 17.8,−4.1, −4.5, −4.9, −5.3; HRMS (ESI-TOF) m/z calcd for C₂₇H₄₄N₃O₇Si₂[M−H]⁻ 578.2718, obsd 578.2718.

6-O-Acetyl-2-azido-3-O-benzoyl-4-tert-butyldimethylsilyl-2-deoxy-β-D-glucopyranosidetrichloroacetimidate (S4)

To a solution of S3 (5.01 g, 8.63 mmol) in 60.0 mL THF at 0° C. wasadded glacial AcOH (0.670 mL) and TBAF (1 M in THF, 10.4 mL). Thereaction mixture was stirred at 0° C. for 2 h, then diluted with 200 mLEt₂O, and washed with brine (3×). The organic layer was dried overMgSO₄, filtered through Celite, and concentrated in vacuo. The cruderesidue was dissolved in CH₂Cl₂ (215 mL) and cooled to 0° C.Trichloroacetonitrile (13.0 mL, 130 mmol) and DBU (130 μL, 0.871 mmol)were added, and the solution was stirred for 2 h at 0° C. The reactionmixture was concentrated in vacuo and purified by silica gel flashchromatography (10:1 hexanes:EtOAc) to afford a 10:1 mixture of α:βanomers (3.80 g, 92%) as a light yellow foam. ¹H NMR (500 MHz; CDCl₃): δ8.34 (s, 1H), 8.08 (d, J=7.0 Hz, 2H), 7.61 (t, J=7.5 Hz, 1H), 7.48 (t,J=7.5 Hz, 2H), 6.52 (d, J=3.6 Hz, 1H), 5.73 (dd, J=10.5, 8.4 Hz, 1H),4.42 (d, J=10.5 Hz, 1H), 4.18-4.10 (m, 2H), 4.05 (t, J=9.0 Hz, 1H), 3.64(dd, J=10.5, 3.5 Hz, 1H), 2.09 (s, 3H), 0.77 (s, 9H), 0.03 (s, 3H),−0.14 (s, 3H); ¹³C NMR (126 MHz; CDCl₃): δ 170.58, 165.50, 160.96,133.61, 130.04, 129.67, 128.65, 94.91, 90.78, 73.20, 72.85, 69.26,62.51, 61.63, 29.83, 25.71, 25.71, 25.69, 25.67, 20.92, 18.04, −3.90,−4.79; HRMS (ESI-TOF): m/z calcd for C₂₃H₃₁Cl₃N₄O₇SiNa [M+Na]⁺ 631.0925,obsd 631.0925.

2-(2-((2S)-Bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)ethanol (S5)

Exo-5-norbornene-2-methanol (4.09 g, 33.0 mmol) was dissolved in THF(60.0 mL) with 4 Å molecular sieves (MS) and stirred at room temperature(rt) for 30 min. The solution was cooled to 0° C., and 95% NaH inmineral oil (1.06 g, 41.2 mmol) was added in portions at 0° C. over 30min. A solution of2-[2-[[(1,1-dimethylethyl)dimethylsilyl]oxy]ethoxy]ethyl-1-methanesulfonate(8.20 g, 27.5 mmol) and 18-crown-6 (1.45 g, 5.49 mmol) in THF (15.0 mL)was added dropwise to this solution, stirred for 2 h at rt, and quenchedwith H₂O until there was no gas formation. The resulting solution wasdiluted with CH₂Cl₂, washed with saturated NaHCO₃, dried over MgSO₄, andconcentrated in vacuo. Purification by silica gel flash chromatography(20:1→5:1 hexanes:EtOAc) afforded the desiredtert-butyldimethylsilyl-protected intermediate (7.21 g) in 67% yield. ¹HNMR (500 MHz; CDCl₃): δ 6.00 (d, J=25 Hz, 2H), 3.71 (t, J=5.5 Hz, 2H),3.60-3.28 (m, 8H), 2.72 (s, 1H), 2.69 (s, 1H), 1.65-1.62 (m, 1H),1.26-1.21 (m, 2H), 1.19-1.14 (m, 1H), 1.05-1.01 (m, 1H), 0.83 (s, 9H),0.00 (s, 6H); ¹³C NMR (125 MHz; CDCl₃): δ 147.1, 86.4, 83.0, 81.1, 80.7,73.1, 55.3, 54.0, 51.9, 49.1, 40.0, 36.3, 28.7, 5.1; HRMS (ESI-TOF): m/zcalcd for C18H35O3Si [M+H]+ 327.2356, obsd 327.2356. The intermediatefrom the previous step (7.21 g, 22.1 mmol) was dissolved in THF (100 mL)and cooled to 0° C. A solution of 1 M TBAF in THF (44.2 mL, 44.2 mmol)was added to the reaction mixture dropwise, and the reaction was stirredfor 2 h at 0° C. The reaction was quenched with H₂O and extracted withEtOAc (3×15.0 mL). The organic layers were combined, dried over MgSO₄,and concentrated in vacuo. Purification by silica gel flashchromatography (5:1→1:1 hexanes:EtOAc) afforded the desired product(4.55 g) in 97% yield. ¹H NMR (500 MHz; CDCl₃): δ 6.11-6.04 (m, 2H),3.72-3.37 (m, 10H), 2.80 (s, 1H), 2.74 (s, 1H), 2.48 (m, 1H), 1.71 (m,1H), 1.34-1.23 (m, 3H), 1.13-1.09 (m, 1H); ¹³C NMR (125 MHz; CDCl₃): δ136.9, 136.8, 76.4, 72.2, 70.7, 70.6, 62.1, 45.2, 43.9, 41.8, 39, 30;HRMS (ESI-TOF): m/z calcd for C₁₂H₂₁O₃ [M+H]⁺ 213.1491, obsd 213.1483.

6-O-Acetyl-2-azido-3-O-benzoyl-4-O-tert-butyldimethylsilyl-1-O-(2-(2-((2S)-bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)ethyl)-2-deoxy-β-D-glucopyranoside(S6)

Compounds S4 (444 mg, 0.728 mmol) and S5 (230 mg, 1.08 mmol) wereazeotroped (3×) with toluene (5.00 mL) and dried for 1 h under highvacuum. The starting materials were dissolved in CH₂Cl₂ (5.00 mL) andmixed with activated 4 Å MS for 20 min at rt. The solution was thencooled to −20° C., and BF₃.OEt₂ (0.720 mL of 0.5 M solution in CH₂Cl₂,0.36 mmol) was added dropwise. After stirring at −20° C. for 30 min, thereaction mixture was warmed to 0° C., quenched with Et₃N, and filteredthrough a pad of Celite. After removal of organic solvents in vacuo, theresidue was purified by silica gel flash chromatography (3:1hexanes:EtOAc) to afford 30 (370 mg) in 84% yield as a white solid. ¹HNMR (500 MHz; CDCl₃): δ 8.04 (dt, J=7.1, 1.4 Hz, 2H), 7.56 (t, 1H), 7.44(t, 2H), 6.04 (ddd, J=26.4, 5.7, 2.9 Hz, 2H), 5.15 (dd, J=10.4, 8.9 Hz,1H), 4.58 (dd, J=8.1, 1.2 Hz, 1H), 4.43 (dd, J=12.0, 2.2 Hz, 1H),4.12-4.10 (m, 1H), 4.00 (dt, J=11.2, 4.4 Hz, 1H), 3.92-3.76 (m, 2H),3.75-3.65 (m, 2H), 3.65-3.44 (m, 7H), 3.34 (td, J=9.3, 4.3 Hz, 1H),2.79-2.69 (m, 2H), 2.01 (s, 3H), 1.29-1.26 (m, 1H), 1.07 (dt, J=11.6,3.9 Hz, 1H), 0.72 (s, 7H), −0.02 (s, 3H), −0.22 (s, 3H); ¹³C NMR (126MHz; CDCl₃): δ 170.76, 165.51, 136.68, 136.61, 133.41, 129.93, 128.53,102.22, 77.41, 77.16, 76.90, 76.11, 75.21, 74.37, 70.71, 70.45, 70.30,69.36, 69.29, 64.51, 62.81, 60.46, 45.05, 43.68, 41.57, 38.77, 29.77,25.61, 21.09, 20.96, 17.87, 14.24, −4.02, −4.88; HRMS (ESI-TOF): m/zcalcd for C₃₃H₅₀N₃O₉Si [M+H]⁺ 660.3311, obsd 660.3297.

6-O-Acetyl-2-azido-3-O-benzoyl-1-O-(2-(2-((2S)-bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)ethyl)-2-deoxy-β-D-glucopyranoside(30)

To a solution of S4 (370 mg, 0.607 mmol) in THF (6.00 mL) and pyridine(6.00 mL) at 0° C. was added HF.pyridine (3.50 mL) dropwise. Thereaction mixture was allowed to warm up to rt and stirred overnight. Thereaction was quenched with a saturated NaHCO₃ solution and diluted withEtOAc. The reaction mixture was extracted with EtOAc and then washedwith saturated NaHCO₃, brine, and 10% CuSO₄, dried over MgSO₄, andconcentrated in vacuo. The resulting residue was purified by silica gelflash chromatography (1:1:2 CH₂Cl₂:hexanes:EtOAc) to furnish compound 30(250 mg) in 83% yield as a white solid. ¹H NMR (500 MHz; CDCl₃): δ 8.06(dt, J=6.8, 1.4 Hz, 2H), 7.64-7.54 (m, 1H), 7.49-7.42 (m, 2H), 6.06(ddd, J=24.1, 5.6, 2.9 Hz, 2H), 5.06 (dd, J=10.3, 9.1 Hz, 1H), 4.57 (d,J=8.0 Hz, 1H), 4.44-4.33 (m, 2H), 4.08-4.00 (m, 1H), 3.84 (ddd, J=10.9,6.1, 4.2 Hz, 1H), 3.75-3.71 (m, 2H), 3.68-3.65 (m, 2H), 3.62-3.55 (m,3H), 3.51 (ddd, J=9.5, 6.2, 3.2 Hz, 1H), 3.36 (td, J=9.2, 4.8 Hz, 1H),2.81-2.69 (m, 2H), 2.09 (s, 3H), 1.69 (tt, J=9.4, 5.6 Hz, 1H), 1.34-1.26(m, 2H), 1.08 (ddd, J=11.7, 4.3, 3.3 Hz, 1H); ¹³C NMR (125 MHz; CDCl₃):δ 171.61, 167.00, 149.51, 136.76, 136.64, 133.76, 130.09, 129.17,128.62, 102.36, 76.16, 76.06, 74.17, 70.77, 70.46, 70.33, 69.53, 69.29,63.90, 63.11, 45.09, 43.73, 41.62, 38.82, 29.82, 20.98; HRMS (ESI-TOF):m/z calcd for C₂₇H₃₆N₃O₉ [M+H]⁺ 546.2446, obsd 546.2453

Methyl 3,4-di-O-benzyl-1,2-methylorthoaceto-α-L-idopyranosiduronate (S8)

Methyl 3-O-benzyl-β-L-idopyranuronate 1,2-(methyl-orthoacetate) (S7,1.60 g, 4.52 mmol) was dissolved in neat benzyl bromide (20.0 mL, 5.00mL/mmol, filtered through a pad of activated basic alumina immediatelybefore use) along with activated 4 Å MS (500 mg/mmol). To this was addedTBAI (1.34 g, 3.62 mmol), and the resulting mixture was stirred at rtfor 15 min. Freshly prepared Ag₂O (4.19 g, 18.1 mmol) was added to themixture and stirred at rt for 8 h the absence of light. The reactionmixture was diluted with Et₂O, filtered through Celite, and concentratedin vacuo. Purification by silica gel flash chromatography (6:1→2:1,hexanes:EtOAc+0.5% Et₃N) afforded the desired product (1.51 g) in 75%yield. ¹H NMR (500 MHz; CDCl₃): δ 7.38-7.19 (m, 10H), 5.50 (d, J=2.9 Hz,1H), 4.58-4.52 (m, 3H), 4.44-4.37 (m, 2H), 4.13-4.08 (m, 1H), 4.08-4.04(m, 1H), 3.85-3.80 (m, 1H), 3.71 (s, 3H), 3.25 (s, 3H), 1.67 (s, 3H);¹³C NMR (125 MHz; CDCl₃): δ 169.31, 137.72, 137.07, 128.74, 128.39,128.36, 127.99, 127.83, 127.81, 124.50, 96.85, 77.41, 77.16, 76.91,76.31, 72.82, 72.77, 72.00, 71.40, 71.19, 52.43, 49.25, 25.08; HRMS(ESI-TOF): m/z calcd for C₂₄H₂₈O₈ [M+Na]⁺ 467.1687, obsd 467.1675.

Methyldibutylphosphate-2-O-acetyl-3,4-di-O-benzyl-α-L-idopyranosiduronate (31)

Compound S8 (688 mg, 1.55 mmol) was azeotroped with toluene (3×10.0 mL)and dried for 1 h under vacuum. Activated 4 Å MS (500 mg/mmol) underflux of argon were added, and the starting material was dissolved inCH₂Cl₂ (1.00 mL/0.100 mmol). After stirring for 10 min at rt, themixture was added dropwise via cannula (within 30 min) to a 3 M solutionof HOPO(OBu)₂ in CH₂Cl₂ (977 μL, 4.65 mmol) in the presence of 4 Å MS(500 mg/mmol HOPO(OBu)₂). After 5 h at rt, the reaction was cooled to 0°C. and Et₃N (865 μl, 6.20 mmol) was added. The solution was warmed to rtand filtered through a pad of deactivated silica gel. The resultingmixture was concentrated in vacuo, and purified by silica gel flashchromatography (4:1→2:1 hexanes:EtOAc+0.5% Et₃N) to afford the desiredproduct (888 mg) in 92% yield. ¹H NMR (500 MHz; CDCl₃): δ 7.45-7.16 (m,10H), 5.86 (d, J=6.9, 1.3 Hz, 1H), 5.03 (dt, J=2.8, 1.2 Hz, 1H), 4.95(d, J=2.5 Hz, 1H), 4.83-4.57 (dd, 2H), 4.55-4.40 (dd, 2H), 4.15-3.98 (m,4H), 3.95-3.82 (m, 2H), 3.74 (s, 3H), 2.04 (s, 3H), 1.69-1.59 (m, 4H),1.46-1.32 (m, 4H), 0.99-0.87 (m, 6H); ¹³C NMR (126 MHz; CDCl₃): δ169.65, 168.74, 137.21, 137.05, 128.30, 128.22, 127.86, 127.82, 127.80,127.66, 95.33, 95.29, 77.42, 77.16, 76.90, 73.49, 72.32, 72.08, 70.62,68.89, 67.82, 67.77, 67.75, 67.71, 66.83, 66.76, 52.04, 32.06, 32.00,31.95, 20.71, 18.44, 13.42, 13.41; HRMS (ESI-TOF): m/z calcd forC₃₁H₄₄O₁₁P [M+H]⁺ 623.2616, obsd 623.2624.

Example 8: Synthesis of Glycopolymers 26

An exemplary synthetic route for glycopolymers is shown in FIG. 11.

Methyl2-O-acetyl-3,4-di-O-benzyl-α-L-idopyranosiduronate-(1→4)-6-O-acetyl-2-azido-3-O-benzoyl-1-O-(2-(2-((2S)bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)ethyl)-2-deoxy-β-D-glucopyranoside(29)

Donor 31 (51.6 mg, 0.0829 mmol) and acceptor 30 (29.7 mg, 0.0592 mmol)were azeotroped with toluene (3×5.00 mL) and dried for 2 h under highvacuum. The mixture was dissolved in CH₂Cl₂ (2 mL/0.100 mmol of 5),cooled to −30° C. for 2 min, and TMSOTf (22.0 μL, 0.0960 mmol) was addeddropwise. After stirring at −30° C. for 1 h, the reaction mixture wasquenched with Et₃N (2 eq) and filtered through a pad of Celite. Afterremoval of organic solvents in vacuo, the residue was purified by silicagel flash chromatography (3:1 hexanes:EtOAc) to afford compound 29 (47.5mg) in 85% as a colorless oil. ¹H NMR (500 MHz; CDCl₃): δ 8.07-8.02 (m,2H), 7.59-7.52 (m, 1H), 7.42 (dd, J=8.3, 7.2 Hz, 2H), 7.36-7.19 (m, 8H),7.12-7.07 (m, 2H), 6.07 (ddd, J=25.9, 5.7, 3.0 Hz, 2H), 5.25 (dd,J=10.3, 9.2 Hz, 1H), 5.09 (d, J=4.7 Hz, 1H), 4.77-4.72 (m, 1H), 4.63 (d,J=12.1 Hz, 1H), 4.57-4.52 (m, 2H), 4.41 (dd, J=12.2, 2.0 Hz, 1H), 4.32(dd, 2H), 4.22-4.15 (m, 2H), 4.08-3.98 (m, 2H), 3.88-3.77 (m, 1H), 3.72(dd, J=5.7, 3.6 Hz, 2H), 3.68-3.50 (m, 6H), 3.41 (s, 3H), 3.39-3.32 (m,1H), 2.82-2.72 (m, 2H), 2.04 (s, 3H), 1.96 (s, 3H), 1.78-1.61 (m, 1H),1.33-1.19 (m, 2H), 1.13-1.06 (m, 1H); ¹³C NMR (125 MHz; CDCl₃): δ170.73, 170.01, 169.66, 165.46, 137.91, 137.31, 136.75, 136.72, 133.09,130.12, 129.90, 128.50, 128.43, 128.32, 128.04, 127.91, 127.81, 127.65,102.39, 99.00, 77.41, 77.16, 76.91, 76.20, 76.09, 75.09, 74.16, 73.03,73.01, 72.77, 72.60, 70.87, 70.74, 70.55, 70.43, 70.41, 70.25, 69.55,64.45, 62.28, 60.52, 51.79, 45.12, 43.75, 41.66, 38.86, 29.84, 21.19,20.97, 20.90, 14.33; HRMS (ESI-TOF): m/z calcd for C₅₀H₅₉N₃O₁₆Na [M+Na]⁺980.3788, obsd 980.3783.

3,4-Di-O-benzyl-α-L-idopyranosiduronate-(1→4)-2-azido-1-O-(2-(2-((2S)bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)ethyl)-2-deoxy-β-D-glucopyranoside(32)

To a solution of 29 (60.0 mg, 0.0636 mmol) in THF (5.00 mL) at −10° C.were added LiOH (1 M solution, 0.636 mL) and H₂O₂ (30% solution, 0.365mL) simultaneously. After stirring for 12 h at rt, the mixture wascooled to −10° C., and MeOH (5.00 mL) and NaOH (4 M solution, 0.790 mL)were added. This reaction mixture was stirred at rt for 12 h,neutralized with Amberlite® IR120 Hydrogen form resin, filtered, andconcentrated in vacuo. The residue was first purified by Sephadex® LH-20chromatography (1:1 CH₂Cl₂:MeOH), followed by silica gel flashchromatography (40:2:1 EtOAc:MeOH:H₂O) to furnish compound 32 (40.2 mg)in 82% as a colorless oil. ¹H NMR (500 MHz; CD₃OD): δ 7.39-7.20 (m,10H), 6.06 (ddd, J=20.5, 5.7, 3.0 Hz, 2H), 4.99 (d, J=3.6 Hz, 1H), 4.69(d, J=11.3 Hz, 1H), 3.71-3.67 (m, 3H), 4.65 (d, J=2.6 Hz, 1H), 4.62-4.54(m, 3H), 4.40 (d, J=8.1 Hz, 1H), 4.09 (ddd, J=3.6, 2.7, 0.8 Hz, 1H),3.99 (dt, J=11.2, 4.3 Hz, 1H), 3.83 (dd, J=12.1, 2.3 Hz, 1H), 3.81-3.69(m, 2H), 3.68-3.56 (m, 6H), 3.52 (dd, J=9.5, 6.3 Hz, 1H), 3.46-3.36 (m,2H), 3.17 (dd, J=9.8, 8.2 Hz, 1H), 2.74 (d, J=27.8 Hz, 2H), 1.65 (dddd,J=14.7, 8.4, 4.4, 1.4 Hz, 1H), 1.36-1.26 (m, 3H), 1.21 (dddd, J=11.5,8.4, 2.4, 0.8 Hz, 1H), 1.16-1.09 (m, 1H); ¹³C NMR (126 MHz; CD₃OD): δ176.22, 139.73, 139.44, 137.69, 137.55, 129.45, 129.29, 128.85, 128.77,128.60, 103.76, 103.04, 79.61, 78.74, 78.61, 77.02, 76.78, 75.10, 74.39,73.31, 72.06, 71.62, 71.56, 71.40, 70.79, 69.99, 68.38, 61.87, 45.85,44.89, 42.76, 40.08, 30.63; HRMS (ESI-TOF): m/z calcd for C₃₈H₄₉N₃O₁₃Na[M+Na]⁺ 778.3158, obsd 778.3160.

3,4-Di-O-benzyl-2-O-sulfonato-α-L-idopyranosiduronate-(1→4)-2-azido-3,6-di-O-sulfonato-1-O-(2-(2-((2S)bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)ethyl)-2-deoxy-β-D-glucopyranoside (33)

To a solution of 32 (42.8 mg, 0.0567 mmol) in anhydrous DMF (3.50 mL) atrt was added SO₃.NMe₃ (237 mg, 1.70 mmol). The reaction mixture wasstirred at 55° C. for 2 d and then quenched with Et₃N (1.00 mL) and MeOH(3.00 mL). After concentration in vacuo, the residue was purified bySephadex® LH-20 (1:1 CH₂Cl₂:MeOH) to obtain compound 33 (39.7 mg) in 71%yield as a white solid. ¹H NMR (500 MHz; CD₃OD): δ 7.53-7.45 (m, 2H),7.42-7.20 (m, 8H), 6.08 (ddd, J=23.8, 5.7, 3.0 Hz, 2H), 4.67 (d, J=12.1Hz, 2H), 4.48 (t, J=9.4 Hz, 2H), 4.36 (t, J=9.0 Hz, 1H), 4.31-4.18 (m,4H), 4.00 (dt, J=11.3, 4.4 Hz, 1H), 3.92 (s, 1H), 3.81 (dt, J=10.7, 4.9Hz, 2H), 3.75-3.57 (m, 9H), 3.53 (dt, J=9.5, 6.3 Hz, 1H), 3.41 (td,J=9.2, 5.8 Hz, 2H), 2.75 (d, J=30.2 Hz, 2H), 1.67 (dddd, J=9.4, 7.5,5.3, 3.7 Hz, 1H), 1.40-1.32 (m, 3H), 1.23 (ddd, J=11.0, 8.3, 2.3 Hz,1H), 1.14 (dt, J=11.7, 3.9 Hz, 1H); ¹³C NMR (126 MHz; CD₃OD): δ 179.50,157.52, 153.60, 144.04, 137.60, 129.40, 128.68, 127.75, 112.94, 110.71,106.19, 101.94, 79.98, 77.03, 71.56, 70.36, 67.22, 63.39, 56.82, 45.87,44.90, 42.77, 40.04, 38.10, 35.00, 30.66, 30.65, 21.09, 20.04, 10.98;HRMS (ESI-TOF): m/z calcd for C₃₈H₄₈N₃O₂₂S₃Na [M+Na]⁻ 1017.1795, obsd1017.9629.

3,4-Di-O-benzyl-2-O-sulfonato-α-L-idopyranosiduronate-(1→4)-2-sulfonatamido-3,6-di-O-sulfonato-1-O-(2-(2-((2S)bicyclo[2.2.1]hept-5-en-2-ylmethoxy)ethoxy)ethyl)-2-deoxy-β-D-glucopyranoside(28)

To a solution of 33 (78.0 mg, 0.079 mol) in THF (10.0 mL) and NaOH (0.1M solution, 4.70 mL) at rt was added PMe₃ (1 M in THF, 0.630 mL). Thereaction mixture was stirred overnight and then neutralized with a 0.1 Msolution of HCl. After concentration in vacuo, the residue was purifiedby Sephadex® LH-20 (MeOH), and the crude product was used for the nextreaction. ¹H NMR (500 MHz; CD₃OD): δ 7.47 (d, J=7.8 Hz, 2H), 7.43-7.20(m, 6H), 6.09 (ddd, J=22.5, 5.7, 2.9 Hz, 2H), 4.65 (d, J=11.8 Hz, 2H),4.56 (d, J=10.3 Hz, 2H), 4.33 (q, J=8.2 Hz, 3H), 4.25 (s, 1H), 4.08-3.98(m, 1H), 3.94 (s, 2H), 3.87-3.77 (m, 1H), 3.77-3.58 (m, 13H), 3.59-3.49(m, 1H), 3.48-3.39 (m, 1H), 2.76 (d, J=37.3 Hz, 2H), 1.73-1.65 (m, 1H),1.36 (dd, J=20.9, 12.3 Hz, 3H), 1.29-1.20 (m, 1H), 1.21-1.11 (m, 1H);HRMS (ESI-TOF): m/z calcd for C₃₈H₄₉NO₂₂S₃Na [M+2H+Na]⁻ 990.1806, obsd990.2563. To a solution of this intermediate (78.2 mg, 0.079 mmol) inanhydrous pyridine (15 mL) and Et₃N (3 mL) at rt was added SO₃.pyr (375mg, 2.37 mmol). The reaction mixture was stirred for 24 h and thenquenched with Et₃N (5 mL) and MeOH (10 mL). After concentration invacuo, the residue was purified by Sephadex® LH-20 (MeOH) to obtain 28(45.5 mg) in 54% yield over two steps as a white solid. ¹H NMR (500 MHz;CD₃OD): δ 7.58-7.51 (m, 2H), 7.41-7.20 (m, 8H), 6.10 (ddd, J=26.5, 5.8,3.0 Hz, 2H), 4.97 (d, J=12.4 Hz, 1H), 4.72 (d, J=12.4 Hz, 1H), 4.66 (d,J=6.3 Hz, 1H), 4.49 (d, J=12.3 Hz, 2H), 4.43 (d, J=10.8 Hz, 1H),4.33-4.18 (m, 4H), 4.18-4.09 (m, 2H), 4.03 (ddd, J=9.9, 5.8, 3.8 Hz,3H), 3.90-3.61 (m, 7H), 3.57 (td, J=9.9, 6.4 Hz, 1H), 3.52-3.39 (m, 1H),3.37 (s, 1H), 2.77 (d, J=20.4 Hz, 2H), 1.72 (dt, J=13.6, 6.8 Hz, 1H),1.39-1.29 (m, 3H), 1.25 (d, J=7.1 Hz, 1H), 1.19-1.14 (m, 1H); ¹³C NMR(126 MHz; (CD₃)₂SO): δ 172.59, 138.60, 138.46, 136.53, 136.35, 128.33,128.01, 127.82, 127.65, 127.32, 126.96, 101.02, 76.54, 74.88, 73.70,71.30, 70.88, 70.36, 70.05, 69.81, 69.63, 69.50, 68.97, 67.32, 64.83,55.73, 55.31, 44.67, 43.20, 41.00, 38.46, 31.28, 29.26, 22.09, 21.01,18.65, 13.96; HRMS (ESI-TOF): m/z calcd for C₃₈H₄₆NO₂₅S₄Na [M+Na]⁻1067.1134, obsd 1067.7520.

Polymerization Procedure:

In a typical polymerization experiment, a small vial was charged withmonomer 28 (22.0 mg, 0.021 mmol) and a small stir bar under the flow ofargon. To this was added dry, degassed MeOH (150 μL) and dry, degassed(CH₂Cl)₂ (750 μL). The ratio of MeOH:(CH₂Cl)₂ varied from 1:4 to 1:2.5depending on the target polymer length. A ratio of 1:4 MeOH:(CH₂Cl)₂ wasused to synthesize polymers 35-4, 35-6, and 35-8, a ratio of 1:3MeOH:(CH₂Cl)₂ was used for polymers 35-10 and 35-15, and a ratio of1:2.5 MeOH:(CH₂Cl)₂ was used for the synthesis of polymers 35-30 and35-45. Increased amounts of MeOH were required to accommodate theinsolubility of the longer polymer chains. The monomer solution washeated to 55° C., stirred for 10 min, and [(H₂IMes)(py)₂(Cl)₂Ru═CHPh](34, 0.013 M stock solution in (CH₂Cl)₂) was quickly added via syringe.The solution was stirred at 55° C. for 2 h until it became cloudy andTLC of the reaction mixture indicated complete consumption of themonomer. The polymerization reaction was quenched by the addition ofethyl vinyl ether (300 μL). The solvent was removed in vacuo to obtain asolid precipitate, which was dissolved in a minimal amount of 10:1CH₂Cl₂:MeOH and slowly added to 25 mL of hexanes in a 50 mL beaker. Tocollect the precipitant, this solution was centrifuged, and the hexaneslayer was decanted to obtain polymer 35 as a white solid. ¹H NMR of thecrude product showed disappearance of the norbornene olefinic protons at˜6 ppm, indicating completion of the polymerization reaction. Theresulting pellet was then dried and purified using a Sephadex G-50column eluted with H₂O.

The series of purified polymer 35 was characterized by ¹H NMRspectroscopy and gel permeation chromatography (GPC)/size exclusionchromatography-multi-angle light scattering (SEC-MALS). For GPCexperiments, the polymers were dissolved in a solution of 0.2 M LiBr inDMF and analyzed on two I-series Mixed Bed Low Molecular Weight ViscoGelcolumns (Viscotek), connected in series with a DAWN EOS MALS detectorand an Optilab DSP differential refractometer (both from WyattTechnology). ¹H NMR (400 MHz; D₂O): δ 7.39-7.11 (m, 10H), 4.94-4.87 (m,1H), 4.81-4.68 (m, 1H), 4.69-4.60 (m, 1H), 4.61-4.53 (m, 1H), 4.53-4.44(m, 2H), 4.38 (bs, 3H), 4.28 (bs, 2H), 4.23-4.09 (m, 3H), 4.10-4.00 (m,1H), 4.00-3.74 (m, 4H), 3.73-3.37 (m, 6H), 4.23-4.09 (m, 2H), 2.22-1.99(m, 2H), 1.99-1.70 (m, 3H), 1.69-1.37 (m, 3H).

Polymer Hydrogenolysis:

Polymer 35 (21 mg) and 20% Pd(OH)₂/C (126 mg, 6× by weight) weredissolved in a 1:3 mixture of phosphate buffer (80 mM, pH=7.2) and MeOH(2 mL). The reaction vessel was equipped with a H₂ balloon and stirredat rt for 2 days. The reaction mixture was filtered through a Milliporenylon membrane (pore size 0.45 μM, filter diameter 47 mm, Product#HNWPO4700), and the membrane filter was washed with warm water (37°C.). The filtrate was lyophilized, dissolved in H₂O (500 μL), andpurified through a Sephadex G-50 column eluted with H₂O. The resultinglyophilized product was then desalted using a Sephadex G-25 column inH₂O. Product fractions were lyophilized to obtain the target polymer 26.¹H NMR (500 MHz; D₂O): δ 5.16 (bs, 1H), 4.85-4.73 (m, 1H), 4.63-4.52 (m,1H), 4.40-4.32 (m, 1H), 4.31-4.20 (m, 3H), 4.05 (bs, 1H), 4.03-3.92 (bs,3H), 3.82-3.53 (m, 16H), 3.51-3.44 (m, 1H), 3.37-3.22 (bs, 2H),1.89-1.70 (m, 2H), 1.68-1.55 (bs, 1H), 1.51-1.08 (m, 8H).

Properties of synthesized polymers are listed in Table 4:

TABLE 4 Heparin Glycopolymer Properties Mol % n M_(n) Entry Polymer 34MeOH:(CH₂Cl)₂ (DP) (g/mol) PDI 1 35-4  30 1:4 4 4,373 2.03 2 35-6  171:4 6 6,167 1.25 3 35-10 10.7 1:3 10 11,207 1.29 4 35-15 6.5 1:3 1515,452 1.32 5 35-30 5.2   1:2.5 30 32,721 1.41 6 35-45 2.0   1:2.5 4542,970 1.25 7 27-35 4.0  1:10 155 164,345 1.62wherein DP, number average molecular weight (Mn), and polydispersityindex (PDI) were determined by SEC-MALS in 0.2 M LiBr in DMF or 100 mMNaNO₃, 200 ppm NaN₃ in H₂O.

Example 9: Chromogenic Assays for the Measurement of Anti-FXa andAnti-FIIa Activity

All reagents were prepared according to the manufacturer's instructionsand incubated at 37° C. for 15 min. Varying concentrations of heparin,low molecular weight heparin, Arixtra® or glycopolymers (0.0005-500μg/mL; 40 μL) and ATIII (0.04 IU; 40 μL) were added to a microcentrifugetube, mixed, and incubated at 37° C. for 2 min. To this, FXa (0.32 μg;40 μL) was added and was incubated at 37° C. for exactly 2 min (stage1), then FXa chromogenic substrate (48 μmol; 40 μL) was added. Exactly 2min later (stage 2), the reaction was stopped by introducing citric acid(240 μL; 20 g/L solution). Absorbance at 405 nm was measured on a UvikonXL spectrophotometer. The sample blank was obtained by mixing thereagents in reverse order from that of the test, i.e. citric acid, FXasubstrate, FXa, ATIII, and heparinized sample. The sample blank valuewas deducted from the absorbance measured for the corresponding assay.

All reagents were prepared according to the manufacturer's instructionsand incubated at 37° C. for 15 min. Varying concentrations of heparin,low molecular weight heparin, or glycopolymers (0.0005-500 μg/mL; 40 μL)and ATIII (0.01 IU; 40 μL) were added to a microcentrifuge tube, mixed,and incubated at 37° C. for 2 min. To this, FIIa (1.2 nkat; 40 μL) wasadded and was incubated at 37° C. for exactly 2 min (stage 1), then FIIachromogenic substrate (0.05 mmol; 40 μL) was added. Exactly 2 min later(stage 2), the reaction was stopped by introducing citric acid (240 μL;20 g/L). Absorbance at 405 nm was measured. The sample blank wasobtained by mixing the reagents in reverse order from that of the test,i.e. citric acid, FXa substrate, FXa, ATIII, and heparinized sample. Thesample blank value was deducted from the absorbance measured for thecorresponding assay.

The anticoagulant activities of the glycopolymers, heparin, LMW heparin(LMWH) and Arixtra were assessed and compared by the above-mentionedassays, with the results shown in FIG. 12 and Table 5.

TABLE 5 Biological Activity of Heparan Sulfate (HS) GlycopolymersAnti-FXa Anti-FIIa APTT PT IC50 (nM) IC50 (nM) (s) (s) 26-4 >2000 >200032.5 ± 0.3 13.2 ± 0.1 26-6 >2000 >2000 32.2 ± 0.2 13.2 ± 0.326-10 >2000 >2000 59.6 ± 0.3 15.8 ± 0.4 26-15 1470 ± 578 >2000 82.9 ±0.6 23.4 ± 1.9 26-30 684 ± 60 577 ± 31 100.8 ± 0.6  50.8 ± 6.3 26-45 5.76 ± 0.04 0.114 ± 10⁻⁴ 119.4 ± 0.5  52.2 ± 7.8 27-35 >2000 >2000 46.1± 0.4 12.7 ± 0.1 None >2000 >2000 31.2 ± 0.3 13.3 ± 0.1 Heparin 16.5 ±1.2 11.0 ± 0.1 >180  84.2 ± 17.8 LMWH 526 ± 71 >2000 117 ± 3  14.8 ± 0.2Arixtra 11.0 ± 0.1 >2000 78.3 ± 0.4 15.1 ± 0.3

LMWH, and Arixtra attenuated FXa activity in the presence of ATIII, withhalf maximal inhibitory concentrations (IC50) of 16.5±1.2, 526±71, and11.0±0.1 nM, respectively (Table 5). Glycopolymer 26-45 showed greateranti-FXa activity compared to the clinical anticoagulants, exhibiting anIC50 value of 5.76±0.04 nM (FIG. 13A, Table 5). The precise contributionfrom the glucosaminyl 3-O-sulfate modification on unit H was alsoassessed by comparing the activity of the glycopolymers to that of3-O-desulfated glycopolymer 27-35. This single alteration in sulfationpattern reduced the anti-FXa activity, reaffirming the importance of3-O-sulfation and the specificity of the polymer interaction with ATIII(FIG. 13A, Table 5).

Also, as shown in FIG. 12, FIG. 13B and Table 5, heparin displayedstrong anti-FIIa activity (IC50=11.0±0.1 nM), whereas LMWH and Arixtrahad no appreciable activity. While the glycopolymer 26-45 was found tobe 100-fold more potent than heparin at inhibiting FIIa (IC50=114±1 pM).

Example 10: Chromogenic Assays for the Measurement of Platelet Factor 4(PF4) Neutralization

All reagents were prepared according to the manufacturer's instructionsand incubated at 37° C. for 15 min. Varying concentrations of heparin,low molecular weight heparin, or glycopolymers (0.0005-500 μg/mL; 40 μL)and ATIII (0.01 IU; 40 μL) were added to a microcentrifuge tube, mixed,and incubated at 37° C. for 2 min. To this, FIIa (1.2 nkat; 40 μL) wasadded and was incubated at 37° C. for exactly 2 min (stage 1) in thepresence or absence of PF4 (20 μg mL-1). FIIa chromogenic substrate(0.05 mmol; 40 μL) was then added and incubated for exactly 2 min (stage2). The reaction was stopped by introducing citric acid (240 μL), andabsorbance at 405 nm was measured. The sample blank was obtained bymixing the reagents in reverse order from that of the test, i.e. citricacid, FXa substrate, FXa, ATIII, and heparinized sample. The sampleblank value was deducted from the absorbance measured for thecorresponding assay.

To assess potential interaction with PF4, the ability of PF4 toneutralize the anti-FIIa activity of glycopolymers 26-45 and 26-30 wasevaluated. PF4 was added to the glycopolymers or heparin (0.5-500 μg/mL)in the presence of ATIII and excess FIIa, and FIIa activity was measuredusing the same chromogenic assay. Both heparin (FIG. 14; A) andglycopolymer 26-45 (FIG. 14; C) interacted strongly with PF4, theanti-FIIa activity of glycopolymer 1-30 was partially neutralized by PF4(FIG. 14; B), indicating PF4 reactivity associated with HIT can beminimized by modulating the polymer length.

Example 11: Activated Partial Thromboplastin Time and Prothrombin TimeAnalysis

Plasma/anticoagulant samples were prepared by mixing 300 μL of theheparin standard or glycopolymer (150 μg/mL) in 0.9% saline and 2.7 mLof citrated human plasma. The tube was inverted 3 times to mix thesample thoroughly. Samples were analyzed by the UCLA Clinical &Translational Research Laboratory using a Sysmex® CA-1500 CoagulationAnalyzer (Siemens AG, Erlangen, Germany). Clotting time in the absenceof an anticoagulant was determined using 0.9% saline solution water (300μl). Each clotting assay was performed in triplicate.

Samples were prepared by mixing 300 μL of the heparin standard orglycopolymer (150 μg/mL) in 0.9% saline with 2.7 mL citrated humanplasma. The tube was inverted 3 times to mix the sample thoroughly.Samples were analyzed by the UCLA Clinical & Translational ResearchLaboratory using a Sysmex® CA-1500 Coagulation Analyzer (Siemens AG,Erlangen, Germany). Clotting time in the absence of an anticoagulant wasdetermined using 0.9% saline solution water (300 μL). Each clottingassay was performed in triplicate.

The activated partial thromboplastin time (APTT) and prothrombin time(PT) of each compound were measured to determine whether the intrinsicand/or extrinsic pathways of the blood coagulation cascade,respectively, were inhibited. As shown in Table 5, heparin increasedboth the APTT and PT for clotting compared to the saline control,whereas LMWH and Arixtra at the same concentration increased only theAPTT. Notably, the APTT of the glycopolymers could be controlled byvarying the polymer length, with a minimum of 10 disaccharide epitopes(26-10) required to prolong the APTT. A slight increase to 15 units(26-15) endowed the polymer with APTT properties similar to Arixtra,whereas the PT was not appreciably altered in either case. Glycopolymers26-30 and 26-45 modulated both the APTT and PT, in agreement with theirability to inhibit FXa and FIIa in vitro. While the APTTs of 26-30 and26-45 were comparable to those of LMWH and Arixtra, their PTs moreclosely resembled that of heparin.

While preferred embodiments of the present invention have been shown anddescribed herein, it will be obvious to those skilled in the art thatsuch embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the invention. It should be understoodthat various alternatives to the embodiments of the invention describedherein may be employed in practicing the invention. It is intended thatthe following claims define the scope of the invention and that methodsand structures within the scope of these claims and their equivalents becovered thereby.

What is claimed is:
 1. A glycopolymer comprising a plurality ofrepeating units, wherein each of said repeating units comprises asaccharide moiety (SA), a linking group (L) and a polymer backbonemoiety (PB), wherein said repeating units are optionally connected byone or more carbon-carbon double bonds, wherein said glycopolymer is ofthe formula:

or a pharmaceutically acceptable salt thereof, wherein: n is about 35;R′ and R″ are each independently selected from the group consisting ofunsubstituted or substituted alkyl, unsubstituted or substitutedheteroalkyl, unsubstituted or substituted aryl, unsubstituted orsubstituted heteroaryl, unsubstituted or substituted cycloalkyl, andunsubstituted or substituted heterocycloalkyl; each of the repeatingunit is of the formula:

R₁ and R₄ are sulfate, and R₂ is sulfite; and m is
 2. 2. Theglycopolymer of claim 1, wherein the polydispersity of said glycopolymeris less than about
 4. 3. The glycopolymer of claim 1, wherein saidglycopolymer lacks anti-coagulant activity.
 4. A composition comprisinga substantially homogeneous population of a glycopolymer of claim
 1. 5.A pharmaceutical composition comprising a glycopolymer of claim 1, and apharmaceutically acceptable carrier.
 6. A substrate, immobilized thereona glycopolymer of claim
 1. 7. The glycopolymer of claim 1, wherein R′ isunsubstituted or substituted cycloalkyl.
 8. The glycopolymer of claim 7,wherein R′ is unsubstituted cycloalkyl.
 9. The glycopolymer of claim 1,wherein R″ is unsubstituted or substituted alkyl.
 10. The glycopolymerof claim 9, wherein R″ is unsubstituted alkyl.
 11. The glycopolymer ofclaim 2, herein the polydispersity index of said glycopolymer is lessthan about 1.5.
 12. A pharmaceutical composition comprising aglycopolymer of claim 1, and a second pharmaceutical agent.
 13. A kitcomprising a glycopolymer of claim
 1. 14. The substrate of claim 6,wherein said substrate comprises a solid support.
 15. The substrate ofclaim 6, wherein said substrate is an array.